CN1145641A - 通过随机片段化和重装配进行的dna诱变 - Google Patents

通过随机片段化和重装配进行的dna诱变 Download PDF

Info

Publication number
CN1145641A
CN1145641A CN95191679A CN95191679A CN1145641A CN 1145641 A CN1145641 A CN 1145641A CN 95191679 A CN95191679 A CN 95191679A CN 95191679 A CN95191679 A CN 95191679A CN 1145641 A CN1145641 A CN 1145641A
Authority
CN
China
Prior art keywords
sequence
fragment
polynucleotide
double
heterozygosis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN95191679A
Other languages
English (en)
Inventor
威利姆·P·C·斯泰莫
安德利亚斯·克拉默瑞
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Affymax Technologies NV
Original Assignee
Affymax Technologies NV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=22733354&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=CN1145641(A) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Affymax Technologies NV filed Critical Affymax Technologies NV
Publication of CN1145641A publication Critical patent/CN1145641A/zh
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/545—IL-1
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1027—Mutagenizing nucleic acids by DNA shuffling, e.g. RSR, STEP, RPR
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
    • C12N9/2471—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C12N9/86—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in cyclic amides, e.g. penicillinase (3.5.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6811—Selection methods for production or design of target specific oligonucleotides or binding molecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
    • C12Y305/02—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amides (3.5.2)
    • C12Y305/02006—Beta-lactamase (3.5.2.6)
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C40B40/08—Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)

Abstract

描述了随机片段化后重装配DNA的方法，及其在体外、或体内用于核酸序列突变的应用。具体地讲，描述了生产编码突变蛋白质的核酸片段或多聚核苷酸的方法。本发明也涉及突变、杂合和选择的循环的方法，该方法使蛋白质在体内或体外发生定向分子进化。

Description

通过随机片段化和重装配进行的DNA诱变
                           发明背景
发明领域
本发明涉及生产多核苷酸的方法，所说多核苷酸提供所需的表型和/或编码一种具有可选择的有利的预定特性的蛋白质。一方面，该方法可用于生产和选择编码突变蛋白质的核酸片段。
相关领域的描述
一种生物大分子例如蛋白质，DNA等的活性序列的复杂性已被称为其信息内容(“ IC”；5-9)。一种蛋白质的信息内容定义为活性蛋白对氨基酸的抗性，可以从用于描述一个具有相同功能的相关序列族所需要的不可变氨基酸(bits)的基本数目计数出(9，10)。对随机诱变敏感的蛋白质具有高信息内容。在1974年，制定该定义时，蛋白质的多样性仅表现为分类上的多样性。
分子生物学的发展例如分子文库已使我们可以鉴别更大量的可变碱基，甚至可以从随机文库中选择功能序列。大多数残基可以是变化的，但是通常不都是在同时发生变化，取决于上下文中的补体变化。所以一个含100个氨基酸的蛋白质不仅可能含有2000种不同的突变，而且可能有20100种的突变组合。
信息密度是单位序列长度上的信息含量。酶的活性位点倾向有高的信息密度。相反，酶的可变的连接点有低的信息密度(8)。
在模式文库中目前广泛应用的制造变异蛋白质的方法是易于产生错误的聚合酶链反应(11，12，19)和盒基因诱变(8，20，21，22，40，41，42)，其中用合成的诱变寡核苷酸替代有待最佳化的特定区域。在两种情况下，在原始序列中一些位点周围产生了‘突变云’(4)。
易于产生错误的PCR使用了低精确性的多聚化条件以在一个长序列内导入低水平的随机点突变。利用易于产生错误的PCR对未知序列的片段的混合物进行诱变。但是，计算机模拟显示单独进行点诱变经常由于太慢而不能发生为连续序列演化所需要的模块变化。公开的易于产生错误的PCR方案不允许大于0.5-1.0kb的DNA片段进行增殖，从而限制了其在实际中的应用。此外易于产生错误的重复的循环导致中性突变积聚，这种积聚例如产生一个免疫源蛋白质。
在寡核苷酸定位诱变中，用合成的诱变寡核苷酸取代一个短序列。这种方法不会产生远缘突变的组合，因此不是组合的。相对于巨大的序列长度而言有限的文库大小是指使蛋白质最佳化不可避免地要进行许多个周期的选择。用合成的寡核苷酸进行诱变需要在每次选择周期之后对单个克隆进行测序，随后划分到族，任意地选择单一的族，将它减少到共有序列基元，重新合成该序列基元并重新插入到单个基因中，随后进行另一次选择。该方法构成了统计学上的峡口，该方法劳动强度大并且对于许多个周期的诱变是不实用的。
因此易于产生错误的PCR和寡核苷酸定位诱变适用于对序列进行精细调整的单个周期，当用于多个周期时很快受限制。
利用易于产生错误的PCR对未知序列的片段的混合物进行诱变(11，12)。但是公开的易于产生错误的PCR方案(11，12)遭受了聚合酶的低加工性能。因此，该方案不能导致一个平均大小的基因产生随机诱变。这种无能限制了易于产生错误的PCR的实用性。
易于产生错误的PCR另一个严重的限制是具有序列信息内容的反向突变的生长速率。对于一定的信息内容，文库大小，和诱变速率，反向突变到正向突变的平衡从统计学上阻碍了对进一步改善的选择性(统计学限度)。
最后易于产生错误的重复的循环导致中性突变积聚，这种积聚产生例如免疫原性但是没有结合亲和力。
因此易于产生错误的PCR由于太慢而不能发生为连续序列演化所需要的模块变化(1，2)。
在盒诱变中，通常用(部分)随机化的序列替代单链模板的序列模块。因此从统计学上说可以获得的最大信息内容受到随机序列数(即文库大小)的限制。该方法构成了统计学上的峡口，消除了通常不是最好的其它序列族，但是它具有更长周期的潜力。
此外，用合成的寡核苷酸进行诱变需要在每次选择周期之后对单个克隆进行测序(20)，因此，该方法劳动强度大并且对于许多个周期的诱变是不实用的。
易于产生错误的聚合酶链反应和盒基因诱变最适合于并且被广泛地用于对相对低信息内容区域进行精细加工。一种明显例外的情况是利用易于产生错误的PCR进行许多个周期的扩增和选择可以从随机文库中选择一种RNA连接酶核糖酶(13)。
用于设计重组线性生物学序列例如蛋白质，RNA和DNA的手段并不象已经研制的手段那样有力量已逐渐显而易见。基于在越来越大的文库内研究越来越多的序列而寻找越来越好的突变体，以及增加诱变扩增和选择的循环数是必要的。但是正如上文中讨论的，当用于重复循环时广泛使用的现存的诱变方法有明显的限制。
通过天然的选择和有性的再生产大多数生物体发生演变。有性再生产可确保所选定的个体的后代的基因进行混合和组合。在有丝分裂期间，来自亲代的同源染色体沿其长度方向以互相并列和交叉部分的方式排列，因此交换了遗传材料。DNA进行这样的交换或混合使生物体可以快速地演化(1，2)。在有性重组中，由于在同源环境中所插入的序列具有实用性，因而一旦插入到新的序列中该插入的序列可能仍具有基本信息内容。
Marton等人(27)描述了在体外利用PCR监测具有直接重复序列的质粒中发生的重组。Marton等人公开了由于DNA的断裂或缺口而在PCR期间出现重组。这样产生重组分子。Meyerhans等人(23)也公开了在体外PCR期间DNA重组的出现。
术语应用分子演变(“AME”)是指为特定的有用目的应用演变的设计的规则系统。而对于多核苷酸(3，11-14)，肽和蛋白质(噬菌体(15-17)，lacI(18)和多核糖体已经报道了许多不同的AME的模式文库，在这些模式文库中已经使用随机交叉重组随意地制备重组文库。
从理论上说，在一个100个氨基酸的蛋白质中存在2000个不同的单突变。一个含100个氨基酸的蛋白质可能有20100种的突变组合，该数目太大以致于不能用常规方法彻底地开发。研制一种使所有的这些可能的突变组合产生和对此筛选的系统是有利的。
Winter和其合作者(43，44)已经利用一个体外位点特异性重组系统将用于在噬菌体系统中表达的轻链抗体基因与重链抗体基因进行组合。但是，它们的系统依赖于重组的特定位点并且因而受到限制。Hayashi等人(48)报道了通过重叠延伸和PCR而在单链抗体中(scFv)发生的抗体CDR区域的同步诱变。
Caren等人(45)描述了利用体内随机重组而产生大批量的多位点突变的方法。但是这些方法需要两个不同的质粒文库发生重组，每个文库具有不同的选择性标记。因此这种方法仅限制在相当于所存在的选择性标记数目的有限数的重组，并且这些方法还伴随着使连接到选定的序列上的标记基因数目呈线性增加。
Calogero等人(46)和Galizzi等人(47)报道了在位于质粒上的两个同源但被截短的昆虫毒素基因进行体内重组可产生杂合基因。Radman等人(49)报道了在错配修复酶缺陷的缩主细胞中实质上错配的DNA序列进行体内重组，导致形成杂合分子。
研制一种生产突变蛋白的方法是有利的，利用该方法可以研制易于研究的突变核酸序列的大文库。本文描述的发明涉及点诱变的重复循环的使用，核酸混合以及对高度复杂线性序列例如通过随机重组的蛋白质的体外特定的分子演化的选择。
因此，研制一种用于生产突变DNA，RNA和蛋白质的大量文库以及为所需目的选择特定的突变体的方法是有利的。本文描述的发明涉及诱变的重复循环的使用，体内重组以及对高度复杂线性序列例如通过随机重组的蛋白质，DNA，RNA的体外特定的分子演化的选择。
根据下面的描述和参考附图，本发明的进一步的优点是显而易见的。
                         发明概述
本发明涉及用于制备选定的多核苷酸序列或选定的多核苷酸序列群体的方法，通常所说的序列是被扩增的和/或克隆的多核苷酸形式，从而选定的多核苷酸序列具有用于选择所需要的表型特性(例如，编码多肽，促进被连接的多核苷酸的转录，结合蛋白质等等)。鉴别具有所需结构或功能特性例如结合到预定的生物大分子(例如受体)上的多肽的方法涉及从多肽的大文库中筛选单个的文库成员，该成员具有该多肽的氨基酸所授予的所需的结构和功能特性。
本发明提供了用于生产适合于亲和作用筛选或表型筛选的目的多肽或目的抗体的文库的方法。该方法包括(1)获得第一批大量的选定的文库成员，含有目的多肽或目的抗体和编码所说目的多肽或目的抗体的相关多核苷酸，以及获得所说相关多核苷酸或其复制本，其中所说的相关多核苷酸含有实际上相同序列的一个区域，在所说多核苷酸或复制本中非强制性地引入突变，和(2)在适合于PCR扩增的条件下通常随机地将所说相关多核苷酸或复制本合并和分裂以形成其片段，进行PCR扩增和非强制性地诱变，从而使所说片段同源重组以形成一个重组多核苷酸杂合池(shuffling pool)，从而在第一批大量选定的文库成员中不存在所说杂合池的重组多核苷酸的基本片段(例如，大于10％)，所说杂合池组成了适合于亲和作用筛选的目的多肽或目的抗体的文库。非强制性地，该方法包括从杂合池中筛选文库成员的附加步骤，以便鉴别具有与预定的大分子，例如一种蛋白质受体，肽，寡糖，病毒颗粒，或其它预定的化合物或结构相结合或相互作用(例如催化抗体)的能力的单个杂合文库成员。目的多肽，抗体，模拟肽抗体，和从这样的文库中鉴别出的可变区序列可用于治疗，诊断，研究，和有关目的(例如催化剂，用于提高水溶液渗透性的溶质等等)，和/或可用于一个或多个混合和/或亲和选择的其它的循环。对该方法进行修改，以便使选择步骤除具有与预定的分子亲和结合外还具有表型特性(例如催化活性，稳定性，抗氧化性，抗药性，或授予宿主细胞的可检测表型)。
在一个实施方案中，通过体外PCR将第一批大量选定的文库成员分裂成片段并且同源重组。
在一个实施方案中，体外将第一批大量选定的文库成员分裂成片段，将得到的片段转移到宿主细胞或生物体并且同源重组，体内形成杂合的文库成员。
在一个实施方案中，不将第一批大量选定的文库成员分裂成片段，但是将它克隆在一个脂质体的可复制的载体中或在该载体中扩增为直系重复片段，其中每个重复片段含有一个远缘品种的选定的文库成员序列，将所说载体转移到一个细胞中并且通过内-载体重组方法进行同源重组以便体内形成杂合的文库成员。
在一个实施方案中，提供了体外和体内杂合的组合方法以加强组合多样性。
本发明提供了用于生产适于亲和作用筛选的目的抗体文库的方法。该方法包括(1)获得第一批大量的选定的文库成员，含有目的抗体和编码所说和目的抗体的相关多核苷酸，以及获得所说相关多核苷酸或其复制本，其中所说的相关多核苷酸含有实际上相同的可变区域骨架序列的一个区域，和(2)在适合于PCR扩增的条件下通常随机地将所说相关多核苷酸或复制本合并和分裂以形成其片段，进行PCR扩增和非强制性地诱变，从而使所说片段同源重组以形成一个包含新的CDR组合的重组多核苷酸杂合池，从而在第一批大量选定的文库成员中不存在含有CDR组合的所说杂合池的重组多核苷酸的基本片段(例如，大于10％)，所说杂合池组成了适合于亲和作用筛选的含有CDR预突变的目的抗体的文库。非强制性地，将杂合池用于亲和筛选以选出与预定的表位抗原决定基(抗原)结合的杂合文库成员以及从而选出了大量的选定的杂合文库成员。非强制性地，将选定的杂合文库成员进行杂合并且重复地筛选1-1000个循环或者按照需要直到获得具有所需结合亲和力的文库成员。
因此，本发明的一个方面提供了一种方法，包括将一个或多个突变导入到模板双链多核苷酸中，其中通过将一个或多个单链或双链寡核苷酸加到得到的双链片段群体上，已经将模板双链多核苷酸裂解为所需大小的随机片段，其中所说的寡核苷酸含有一个等同于模板多核苷酸的区域和异源于模板多核苷酸的区域；使得到的双链随机片段和寡核苷酸的混合物变性为单链片段；将得到的单链片段群体与一种聚合酶一起保温，保温条件是使所说的单链片段退火在相当于该单链片段的区域上并且形成诱变的双链多核苷酸；并且按照需要重复上述步骤。
另一方面，本发明涉及生产具有生物学活性的重组蛋白质的方法，包括通过在使所说模板多核苷酸裂解为具有所需大小的随机双链片段的条件下处理含有编码野生型蛋白质的双链模板多核苷酸的样本；将一个或多个单链或双链寡核苷酸加到得到的随机片段群体上，其中所说的寡核苷酸含有一个等同于模板多核苷酸的区域和异源于模板多核苷酸的区域；使得到的双链随机片段和寡核苷酸的混合物变性为单链片段；将得到的单链片段群体与一种聚合酶一起保温，保温条件是使所说的单链片段退火在等同区域上并且形成诱变的双链多核苷酸；按照需要重复上述步骤，以及然后从诱变的双链多核苷酸表达该重组蛋白质。
本发明的第三个方面涉及获得嵌合多核苷酸的方法，包括在使所说模板多核苷酸裂解为具有所需大小的随机双链片段的条件下处理含有不同双链模板多核苷酸的样本，其中所说的不同模板多核苷酸含有一个等同区域和异源区域；使得到的含有被处理样本的双链随机片段变性为单链片段；将得到的单链片段与一种聚合酶一起保温，保温条件是使所说的单链片段退火在等同区域上并且形成一种嵌合的双链多核苷酸序列，该序列含有模板多核苷酸序列；并且按照需要重复上述步骤。
本发明的第四个方面涉及复制模板多核苷酸的方法，包括通过体外将单链模板多核苷酸与由模板多核苷酸的裂解和变性产生的随机单链小片段结合，以及在核酸多聚酶存在下保温所说的核酸片段混合物，保温条件是形成双链模板多核苷酸群体。
本发明还提供了将体外和/或体内多核苷酸杂合方法用于使编码多肽的多核苷酸和/或含有转录调节序列的多核苷酸进行杂合。
本发明还提供了将多核苷酸杂合方法用于使病毒基因(例如衣壳蛋白，针尖糖蛋白，聚合酶，蛋白酶等等)或病毒基因组(例如副粘病毒科，正粘病毒科，疱疹病毒，反录病毒，呼肠病毒，鼻病毒等等)群体进行杂合。在一个实施方案中，本发明提供了使编码整个或部分免疫病毒蛋白的序列进行杂合以产生新组合的表位决定基以及由重组产生的新的表位决定基的方法；这种杂合的病毒蛋白可以含有表位决定基或表位决定基的组合，它们可能在病毒演变(例如流感病毒菌株的重组)的天然环境中产生。
本发明还提供了适于使多核苷酸序列杂合的方法，用于产生基因治疗载体以及复制缺陷型基因治疗构件体，例如可用于人的基因治疗，包括但不限于基于DNA的免疫接种的免疫接种载体，以及抗瘤形成的基因治疗和其它基因治疗方法。
                           附图简述
附图1图解比较由易于产生错误的PCR进行的诱变杂合；(a)起始文库；(b)在亲和选择的第一个周期中选定序列的集合体；(d)选定序列的体外重组(‘杂合’)；(f)在杂合之后亲和选择的第二个周期中选定序列的集合体；(c)易于产生错误的PCR；(e)在易于产生错误的PCR之后亲和选择的第二个周期中选定序列的集合体。
附图2描述了1.0kb LacZα基因片段与10-50bp随机片段的重组。(a)具有LacZα基因的DNA片段经PCR扩增后凝胶电泳的照片。(b)在用DNA酶I(DNAseI)消化之后的DNA片段凝胶电泳下照片。(c)从被消化的LacZα基因DNA片段纯化的10-50bpDNA片段的凝胶电泳的照片；(d)在DNA重组进行指定数目的周期之后10-50bpDNA片段的凝胶电泳的照片；(e)在用引物进行PCR扩增之后重组混合物凝胶电泳的照片。
附图3图解描述了LacZα基因终止密码子突变体和其DNA序列。加方框的区域是异源区，作为标记。该终止密码子定位于较小的框中或者底下划线。“+”表示野生型基因，“-”表示该基因的突变区。
附图4图解描述一个合成的寡核苷酸导入或插入的LacZα基因的重组过程中。
附图5描述了使用大肠杆菌密码子的小鼠IL1-B基因(M)和人IL1-B基因(H)之间同源区域。方框中指出了异源区域。
Figure A9519167900121
表示有二个基因杂合获得的交叉。
附图6图解显示用scFv的抗兔IgG抗体(A10B)的抗体CDR杂合模型系统。
附图7描述在scFv的抗兔IgG抗体(A10B)的杂合DNA中观察到的CDRs的特定组合出现的频率。
附图8描述了在DNA杂合和每个选择周期之后scFv的抗兔IgG抗体的改善的亲和力。
附图9图解描绘了pBR322-Sfi-BL-LA-Sfi和通过直系重复区进行的体内质粒内重组，以及由重新构成一个功能性β-内酰胺酶质粒内重组产生氨苄青霉素抗性菌落的速率。
附图10图解描绘了pBR322-Sfi-2Bla-Sfi和通过直系重复区进行的体内质粒内重组，以及由重新构成一个功能性β-内酰胺酶质粒内重组产生氨苄青霉素抗性菌落的速率。
附图11描述了用于检测将多核苷酸片段导入到产生重组蛋白的细胞之后多个周期同源重组的效率的方法。
附图12图解描绘了通过基因盒在下列位点杂合而产生的载体文库：启动子，前导肽，终止子，可选择的抗药性基因，和复制原点。在每个位点的多条平行线表示对于那个盒具有盒多样性。
附图13图解显示在各个位点适合于通过杂合方法构建原核载体文库的盒的一些例子。
                      优选实施例的描述
本发明涉及用于在随机片段化之后核酸分子发生重组的方法以及将该方法用于DNA序列的诱变。还描述了用于生产编码具有增强的生物活性的突变蛋白的核酸片段的方法。尤其是，本发明还涉及重复循环的诱变，核酸杂合和可以制备具有增强的生物活性的突变蛋白的选择的方法。
本发明涉及产生DNA，RNA或蛋白质突变体的极大文库的方法。该方法在产生可选出所需核酸片段的有关DNA片段方面有特别的优点。特别是本发明还涉及重复循环的诱变，同源重组和可以制备具有增强的生物活性的突变蛋白的选择的方法。
但是在详细讨论本发明之前，首先定义下列术语。
定义
如本文所用，下列术语具有下列含义：
术语“DNA重组”用于指两个相同序列之间出现重组。
相反，术语“DNA杂合”用于指基本上同源但不相同的序列之间的重组，在一些实施方案中，DNA杂合是指非同源重组出现的交叉，例如借助于cre/lox和/或flp/frt系统等等。
术语“扩增”是指增加一个核酸片段的拷贝数。
术语“相同”或“等同”是指两个核酸序列具有相同序列或一个互补序列。因此“等同区域”是指一个核酸片段或多核苷酸的区域相同或互补于另一个多核苷酸或核酸片段。
术语“相应于”是指一个多核苷酸序列与整个或部分参照多核苷酸序列同源(即相同，演化上不严格相关)，或者是指一个多肽序列等同于参照多肽序列。与之相区别，本文中使用的术语“互补于”是指该互补序列与整个或部分参照多核苷酸序列同源。为了描述，核苷酸序列“TATAC”对应于参照序列“TATAC”并且互补于参照序列“GTATA”。
下列术语用于描述两个或多个多核苷酸之间的序列关系：“参照序列”，“比较窗口”，“序列等同性”，“序列等同百分数”和“基本等同”。“参照序列”是指用作为序列比较的基础的一个选定的序列；参照序列可以是较大序列一个子集，例如在序列表给出的全长cDNA或基因序列的片段，例如图1或图2(b)的多核苷酸序列，或者可以包括完整的cDNA或基因序列。通常，参照序列是长度为至少20个核苷酸，通常长度为至少25个核苷酸，以及经常长度为至少50个核苷酸。由于两个多核苷酸可以各(1)含有两个多核苷酸之间相类似的一个序列(即，整个多核苷酸序列的一部分)，(2)进一步可含有两个多核苷酸之间差异较大的一个序列，通常通过借助于“比较窗口”对两个多核苷酸序列进行比较以鉴别和比较定位区序列相似性通常可完成对两个(或多个)多核苷酸序列的比较。
本文中使用的“比较窗口”是指至少20个连接位点片段，其中多核苷酸序列相当于至少20个连接核苷酸的参照序列以及其中将两个序列最佳排列之后与参照序列(不含有叠加或缺失)相比比较窗口中多核苷酸序列的一部分可以含有20％或者更少的叠加或者缺失(即缺口)。采用Smith and Waterman(1981)Adv.App1.Math.2：482的位点同源规则，采用Needleman and Wunsch(1970)J.Mol.Biol.48：443的同源排列规则，根据Pearson and Lipman(1988)Proc.Natl.Acad.Sci.(U.S.A.)85：2444对相似方法的研究，这些规则的计算机应用(GAP，BESTFIT，FASTA，and TFASTA in the Wisconsin Genetic Software Package Release 7.0，GeneticsComputer Group，575 Science Dr.，Madison，WI)，或者通过检查可以完成对比较窗口进行排列而使序列排列最佳，并且选出有各种方法产生的最佳排列(即产生比较窗口的最高同源百分数)。
术语“序列等同性”是指两个多核苷酸序列的比较窗口是相同的(即，以一个核苷酸接一个核苷酸为基础)。术语“序列等同百分数”是通过下列方法计数的：将两个最佳排列的序列的比较窗口进行比较，测定两个序列上出现等同核酸碱基(例如，A，T，C，G，U，或I)的位置的数目以得出错配位置的数目，将错配位置的数目除于比较窗口中总位置数目(即窗口大小)，以及将得到的结果乘于100以产生序列等同百分数。本文中使用的术语“基本等同”是指多核苷酸序列的特性，其中该多核苷酸含有一个序列，在至少20个核苷酸位置，最常见的是至少25-50个核苷酸的比较窗口内，该序列与参照序列至少有80％的序列等同性，优选地至少85％等同以及通常为90-95％序列等同，最通常为99％序列等同，其中通过将参照序列与多核苷酸序列进行比较而计算出序列等同百分数，所说多核苷酸可以含有参照序列比较窗口内总共20％或更少序列的缺失或叠加。
保守的氨基酸组成是指可内部转化的具有相似侧链的残基。例如，具有脂族侧链的一组氨基酸是甘氨酸，丙氨酸，缬氨酸，亮氨酸，和异亮氨酸；具有脂族-羟基侧链的一组氨基酸是丝氨酸，和苏氨酸；具有含酰胺侧链的一组氨基酸是天冬氨酸和谷氨酰胺；具有芳香族侧链的一组氨基酸是苯丙氨酸，酪氨酸和色氨酸；具有碱性侧链的一组氨基酸是赖氨酸，精氨酸和组氨酸；具有含硫侧链第一组氨基酸是半胱氨酸和甲硫氨酸。优选的保守氨基酸组成基团是：缬氨酸-亮氨酸-异亮氨酸，苯丙氨酸-酪氨酸，赖氨酸-精氨酸，丙氨酸-缬氨酸，和天冬氨酸-谷氨酰胺。
术语“同源”是指一个单链核酸序列与一个互补单链核酸序列杂交。杂交程度取决于多个因素，包括序列之间等同数目和杂交条件例如温度和下文中讨论的盐浓度。优选地等同区域是大于约5bp，更优选地等同区域是大于约10bp。
术语“异源”是指一个单链核酸序列不能与另一个单链核酸序列后其互补酸序列杂交。因此，异源区域是指具有不能与另一个核酸或多核苷酸杂交的序列的区域的核酸片段或多核苷酸。例如这样的区域是突变区。
本文中使用的术语“关连”是指品种之间演化和功能上相关的一个基因序列。例如但不限于，在人基因组中，人CD4基因与鼠CD4基因是一个关连基因，因为这两个基因的序列和结构显示它们是高度同源的并且两个基因编码一个蛋白质，该蛋白质通过MHCII型限定的抗原识别而在作为标记的T细胞活化中起作用。
术语“野生型”是指不含有任何突变的核酸片段。一种“野生型”蛋白质是指在天然存在的活性水平上时该蛋白质是有活性的并且含有天然存在的氨基酸序列。
术语“相关的多核苷酸”是指该多核苷酸区域是相同的并且该多核苷酸区域是异源的。
术语“嵌合多核苷酸”是指该多核苷酸含有野生型区域以及被突变的区域。也是指该多核苷酸含有来自一个多核苷酸的野生型区域以及来自另一个相关多核苷酸的野生型区域。
术语“切割”是指用酶消化一个多核苷酸或裂解该多核苷酸。
本文中使用的术语“群体”是指成分例如多核苷酸，核酸片段或蛋白质的集合。一种“混合的群体”是指属于相同核酸或蛋白质族(即相关)但是其序列不同(即不相同)因此其活性不相同的成分的集合。
术语“特定的核酸片段”是指具有一定的端点并且具有一定的核酸序列的核酸片段。两个核酸片段具有两个不同的特定的核酸片段，这两个核酸片段中一个核酸片段具有相同于第二个核酸片段的一部分但是末端不同的序列。
术语“突变”是指野生型核酸序列内发生变化或肽序列内发生变化。这样的突变可以是点突变例如碱基转换或者转变。突变可以是缺失，插入或者复制。
在本文中使用的肽标记中，按照标准用法和规律左手方向是氨基末端方向，右手方向是羧基末端的方向。类似地，除非有特定的说明，单链多核苷酸序列的左手末端是5′末端；双链多核苷酸序列所左手末端是5′末端。5′3′新生RNA转录物的叠加方向是指转录方向；位于具有类似于RNA的序列的DNA链上并且是5′至RNA转录物的5′末端的序列区域称为“上游区域”；位于具有类似于RNA的序列的DNA链上并且是3′至编码RNA转录物的3′末端的序列区域称为“下游区域”。
本文中使用的术语“天然存在”用作为指物体时是指该物体可以在自然界中发现指定事实。例如，存在于可从天然来源分离的生物体(包括病毒)中并且人类并不想在实验室中将它修饰的多肽或多核苷酸序列是天然存在的。通常，术语天然存在是指存在于非病原(未发病)个体中的物体，例如典型物种。
本文中使用的术语“试剂”是指化合物，化合物的混合物，一系列空间定位的化合物(例如，VLSIPS肽列阵，多核苷酸列阵，和/或小分子组合列阵)，生物学大分子，噬菌体肽陈列文库，噬菌体抗体(例如，scFv)陈列文库，多核糖体肽陈列文库，或从生物材料例如细菌，植物，真菌或动物(特别是哺乳动物)细胞或者组织。通过参与下文中描述的筛选方法，评价这些试剂作为抗瘤形成，抗炎性或者apoptosis调节子的活性。通过参与下文中描述的筛选方法，评价这些试剂作为特定蛋白质相互作用抑制剂的活性(即，选择性地抑制两种预定多肽之间的相互作用但是基本上不干扰细胞存活性的一种试剂)。
如本文中使用的，“基本上纯化”是指一个物种以优势种存在(即以摩尔为基础，它在该组合物中比其它单个大分子品种的含量更丰富)，优选地，一种基本纯化组分是指其中该物种的含量至少为存在的所有大分子的50％(以摩尔计)的组合物。通常，一种基本上纯化的组合物含有该组合物中所有大分子的80-90％以上。更优选地，将该物种纯化至基本上匀质(采用常规方法不能检测出该组合物中的污染物)，其中该组合物基本上由单一大分子组成。溶剂，小分子(＜500道尔顿)，基本离子不认为是大分子。
如本文中使用的，术语“生理条件”是指温度，pH，离子强度，粘度，和类似的生化参数，这些参数与活生物体相对应，和/或通常存在于培养的酵母活细胞或哺乳动物细胞内的条件。例如，在常规实验室培养条件下生长的酵母细胞的胞内条件是生理条件。通常用于体外转录的合适的体外反应条件是生理条件。一般地说，体外生理条件包括50-200mMNaCl或KCl，pH 6.5-8.5，20-45℃和0.001-10mM二价阳离子(例如，Mg++，Ca++)；优选地，约150mMNaCl或KCl，pH7.2-7.6，5mM二价离子，通常包括0.01-1.0％非特异蛋白(例如，BSA)。通常存在约0.001-2％的非离子型去污剂(吐温，NP-40，Triton X-100)，典型的是0.05-0.2％(v/v)。由常规方法的实践者选定特定的含水条件。对于常规应用，可以应用下列缓冲含水条件：10-250mMNaCl，5-50mM Tris HCl，pH 5-8，非强制性地加入二价阳离子，和/或金属螯合剂和/或非离子型去污剂和/或膜组分和/或消沫剂和/或闪烁材料。
本文中特定的杂交定义为在第一多核苷酸和第二多核苷酸(例如，具有独特的但是与第一多核苷酸基本相同的序列的多核苷酸)之间形成杂合体，其中在严谨杂交条件下第一多核苷酸优选与第二多核苷酸杂交，其中基本上不相关的多核苷酸序列在混合物中不形成杂合体。
如本文中使用的，术语“单链抗体”是指含有以多肽键连接的VH区和VL区，通常通过间隔肽(例如，[Gly-Gly-Gly-Gly-Ser]x)连接并且在氨基末端和/或羧基末端含有其它的氨基酸序列的一个多肽。例如，单链抗体含有用于连接到编码多核苷酸上的一个限制片段。作为举例，scFv是一个单链抗体。单链抗体通常是由一个或多个多肽片段组成的蛋白质，所说多肽片段至少含基本由免疫球蛋白超家族的基因编码的(例如，参见The Immunoglobulin Gene Superfamily，A.F.Woilliams and A.N.Barclay，in Immunoglogulin Genes，T.Honjo，F.W.Alt，and T.H.Rabbitts，eds.，(1989)Academic Press：San Diego，CA，pp.361-387，引入本文作为参考)，最常见是由啮齿动物，非人灵长类，鸟类，猪，牛，绵羊，山羊，或者人重链或者轻链基因序列编码的10个相邻氨基酸。通常一个单链抗体含有大部分的免疫球蛋白超家族产物以致于保留了与特定的靶分子，通常一种受体或者抗原(表位决定基)结合的特性。
如本文中使用的，术语“互补决定区”和“CDR”是指本领域内认可的术语如由Kabat和Chothia例举的CDR定义，通常也称为极度可变区或极度可变环(Chothia and Lesk(1987)J.Mol.Biol.196：901；Chothia et al.(1989)Nature 342：877；E.A.Kabat et al.，Sequence of Proteins of Immunological Interest(National Institutesof Health，Bethesda，MD)(1987)；and Tramontano et al.(1990)J.Mol.Biol.215：175)。通常可变区域含有天然存在的免疫球蛋白(例如1-110位氨基酸)的约105-115氨基酸氨基末端，尽管更短或更长的可变区域也适合于形成单链抗体。
免疫球蛋白轻链或重链可变区由“主链”区组成，该主链区被三个极度可变区，也称为CDR’s打断。该主链区和CDR’s的长度已被精确地定义(参见“Sequencesof Proteins of Immunological Interest，”E.Kabat et al.，4th Ed.，U.S.Department ofHealth and Human Services Bethesda，MD(1987))。在一个品种内不同轻链或重链的主链区序列是非常保守的。如在本文中使用的，“人骨架区”是一个主链区，它基本上等同(约85％或更多，通常90％-95％或更多)于天然存在的人免疫球蛋白的主链区。作为组成型轻链和重链的组合主链区的一种抗体的主链区可用作为定位和排列CDR’s。该CDR’s主要具有结合到抗原的表位决定基上的作用。
如本文中使用的，术语“可变片段”是指新生肽的一部分，该肽含有一个随机的，假随机的，或限定的核心序列。可变片段可以含有变异的和非变异的残基位置，可以限制一个变异残基位置上残基变异的程度；两种选择由实践者的意见来决定。通常，可变片段的长度是约5-20个氨基酸残基(例如8-10)，但是可变片段也可以更长并且可含有抗体部分或受体蛋白，例如抗体片段，核酸结合蛋白，受体蛋白等等。
如本文中使用的，“随机肽序列”是指由两个或多个氨基酸单体组成并且由随机过程构建的氨基酸序列。一种随机肽可包括主链或支架基元，它可以含有非变异序列。
如本文中使用的，“随机肽文库”是指编码一组随机肽的一组多核苷酸序列，并且指由这些多核苷酸序列编码的该组随机肽，以及含有这些随机肽的融合蛋白。
如本文中使用的，术语“假随机”是指具有有限的可变性的一组序列，以至于例如一个位置上的残基变异程度不同于另一个位置上的残基变异程度，但是任何假随机位置都被允许有一定的残基变异程度，但变异程度是有限的。
如本文中使用的，术语“限定的序列骨架”是指以非随机方法，通常以试验数据和结构资料为基础选出的一组限定的序列；例如限定的序列骨架含有被预测为可形成β-片层结构的一组氨基酸序列或者含有一个亮氨酸拉链型七个一套的重复基元，一个锌--指状物区域，其它为变异区。“限定的序列核心”是变异性有限的一套序列。(1)一个20个常规氨基酸的序列的完全随机的十聚体可以是(20)10序列，(2)一个20个常规氨基酸的序列的假随机的十聚体可以是(20)10序列，但是在特定位置和/或整个序列显示偏向于特定残基，(3)限定的序列核心是一组序列，如果每个残基位置允许是所说20个常规氨基酸(和/或允许的非常规氨基/亚氨基酸)的任一种则该组序列数低于可能序列数的最大数目。通常限定的序列核心在选定单个文库成员序列的片段上或整个长度内含有变异和非变异残基位点和/或含有变异残基位点，这些位点可含有选自于限定的氨基酸残基亚组的一个残基等等。选定的序列核心可以指氨基酸序列或多核苷酸序列。为了说明而不是为了限制，对序列(NNK)10和(NNM)10选定了序列核心，其中N代表A，T，G，或C；K代表G或T，和M代表A或C。
如本文中使用的，“表位决定基”是指一个抗原或其它大分子上能够形成结合作用的组分，它与一种抗体的可变区结合位置发生相互作用。通常这样的结合作用显示为与CDR的一个或多个氨基酸残基发生分子内接触。
如本文中使用的，“受体”是指与一个给定的配体有亲和力的分子。受体可以是天然存在或合成的分子。可以使用非变化状态的受体或者与其它种的结合体形式的受体。可以直接或借助于一个特定的结合物质将受体共价或非共价吸附到结合成员上。受体的例子包括但不限于与特定的抗原决定基(例如病毒上的，细胞，或其它材料)发生反应的抗体，包括单克隆抗体和抗血清，细胞膜受体，碳水复合物和糖蛋白，酶和激素受体。
如本文中使用的，“配体”是指由特定的受体识别的一种分子，例如随机肽或变异片段序列。如本领域内任一熟练技术人员将认识到，一个分子(分子大分子复合物)即可以是受体又可以是配体。通常具有较小分子量的结合配对体被称为配体，具有较大分子量的结合配对体被称为受体。
如本文中使用的，“连接头”或“间隔区”是指分子或连接两个分子的分子团，例如DNA结合蛋白和一种随机肽，并且将两个分子以优选的构型放置，(例如使该随机肽结合到与DNA结合蛋白有基本空间障碍的一个受体上)的一个分子或一组分子。
如本文中使用的，术语“可操作连接”是指多核苷酸元素以一定的功能关系进行连接。当置于功能关系中时一种核酸与另一种核酸序列可操作连接。例如如果实现了该编码序列的转录则一种启动子或增强子可操作连接到一编码序列上。可操作连接是指通常被连接的DNA序列是相邻的，并且如果必须要连接二个蛋白质编码区，它们是相邻的并且在阅读框架内。
方法论
核酸杂合方法是用于核酸片段或多核苷酸集合体的体外或体内同源重组的方法。将相关核酸序列或多核苷酸序列的混合物随机地裂解成片段，并且重新装配以产生重组核酸分子或多核苷酸的文库或混合群体。
与盒诱变相反，单独的杂合和易于产生错误的PCR可以将序列集合体盲目地突变(除引物外没有序列信息)。
最好是用抗体遗传工程的实例来解释在重复选择时使用本发明的诱变杂合比仅用易于产生错误的PCR具有优点。在附图1中显示了下文中描述的DNA杂合方法。起始文库由各种来源的相关序列(即来自天然mRNA的抗体)组成或者由任何类型的单抗体基因诱变(包括杂合)产生。在第一轮亲和选择之后得到了选定的互补决定区的集合(“CDRs”)(图1)。在该图中浓的CDRs使抗体分子与抗体的亲和力增强。杂合方法使所有的CDR1s和所有的CDR2s和所有的CDR3s等等自由组合(图1)。
该方法不同于PCR，不同之处在于它是一个逆转链反应。在PCR中，聚合酶起始位点的数目和分子数目呈指数增长。但是聚合酶起始位点的序列和该分子的序列基本保持相同。相反在随机片段的核酸重组或杂合方法中，起始位点的数目和随机片段的数目(但不是大小)同时降低。对于从整个质粒衍生的片段理论上的关键点是一种单链，大多联体分子。
由于在同源区域出现了交叉，因此首先在相同序列族成员之间出现重组。这阻碍了严重不相容的(例如抗相同抗原的不同表位决定基)的CDRs的组合。观察到在相同反应中多个序列族可以杂合。此外杂合保存了相对位置序列，以至于例如在CDR2位置上未发现CDR1。
稀有的杂合体含有大量的最好的(例如最高亲和力)CDRs并且基于其优异的亲和力可以选出这些稀少的杂合体(图1)。
以高达1006不同方法可以将来自100个不同的选定的抗体序列的集合体的CDRs排列。在DNA序列单个文库中不能体现这大量的排列。因此，根据序列长度和所需序列的多样性决定需要进行的DNA杂合和选择的多次循环。
相反易于产生错误的PCR在同样的相关序列内保留了所有选定的CDRs(图1)，产生了更小的突变云。
在本发明方法中使用的模板多核苷酸可以是DNA或RNA。根据重组或重装配的基因或DNA片段的大小模板具有不同的长度。优选地模板多核苷酸大小从50bp--50kb。在本发明的方法中可以使用含有编码所需蛋白质的所需核酸的完整载体，事实上已经成功地使用。
通过利用PCR反应进行扩增(美国专利№4683201和4683195)或其它扩增或克隆方法可以获得模板多核苷酸。但是，在分裂成片段之前从PCR产物中除去游离引物可以获得更有效的结果。不能合适地除去引物可以导致产生低频率的交叉克隆。
通常模板多核苷酸应该是双链的。要求双链核酸分子可以确保得到的单链核酸片段的区域互相互补，由此可以杂交以形成双链分子。
在该步骤中将具有等同于模板多核苷酸的区域和异源于模板多核苷酸的区域的单链或双链核酸片段加到模板多核苷酸上。还涉及在该步骤中将两个不同但相关的多核苷酸模板混合。
将双链多核苷酸模板和任何增加的双链或单链片段随机地消化为约5bp--5kb或更大的片段。优选地随机片段的大小为约10bp--1000bp，更优选地随机片段的大小为约20bp--500bp。
另一种可选的方法，还涉及在本发明的方法中使用具有多个缺口的双链核酸。一个缺口是在双链核酸的一条链上有一断裂。所说缺口之间的距离优选的是5bp--5kp，更优选的是10bp--1000bp。
可用许多不同的方法消化核酸片段。可用核酸酶，例如DNA酶I或RNA酶消化核酸片段。采用超声处理方法或通过一个有小洞的管可以随机的剪切核酸。
还涉及用一种或多种限制性酶部分消化核酸，以至于从统计学上保留了交叉点。
任何一种特定核酸片段的浓度不会大于总核酸重量的百分之一，更优选的任何一种特定核酸片段的浓度不会大于总核酸重量的0.1％。
在该混合物中不同的特定核酸片段的数目至少为约100，优选的至少约500，更优选至少约1000。
在该步骤中将合成的或天然的单链或双链核酸片段加到随机双链片段上以便提高该核酸片段混合物的不均匀性。
在该步骤还涉及将随机断裂的双链核酸片段群体混合或组合。
如果需要在模板多核苷酸中插入突变，则以相当于总核酸重量20倍的过量加入具有等同于模板多核苷酸的区域和具有异源于模板多核苷酸的区域的单链或双链核酸片段，更优选地，加入相当于总核酸重量10倍的过量的单链核酸片段。
如果需要不同但相关的模板多核苷酸的混合物，以低于1∶100的比例，更优选地以低于1∶40的比例将来自每个模板的核酸片段群体组合。例如，按照需要将野生型多核苷酸与突变多核苷酸群体回交以去除中性突变(例如，产生所选定的表型特性发生非本质变化的突变)。在一个实施例中，加到随机消化的突变多核苷酸上的随机消化的野生型多核苷酸片段的比例是约1∶1-约100∶1，并且更优选的是从1∶1-40∶1。
将随机发生片段的混合群体变性以形成单链核酸片段并且随后重退火。只有具有与其它单链核酸片段同源的区域的那些单链核酸片段将会退火。
通过加热，随机核酸片段可以变性。本领域内技术人员可以确定使双链核酸完全变性所必须的条件。优选地温度是80℃-100℃，更优选地温度是90℃-96℃。可用于使核酸片段变性的其它方法包括压力(36)和pH。
通过冷却可将核酸片段重退火。优选地温度是20℃-75℃，更优选地温度是40℃-65℃。如果基于仅四个基本碱基同源性的平均值需要高频率的交叉，通过使用低退火温度可以加强重组，尽管该方法变为更复杂。出现变性的程度将依赖于单链核酸片段群体之间同源性的程度。
通过加入聚乙二醇(“PEG”)或盐可以加速变性恢复。优选的盐浓度是从0mM-200mM，更优选的盐浓度是从10mM-100mM。所说盐可以是KCl或NaCl。优选地PEG浓度是0％-20％，更优选地5％-10％。
接着在核酸聚合酶和dNTP′s(即dATP，dCTP，dGTP和dTTP)存在下保温退火的核酸片段。核酸聚合酶可以是Klenow片段，Taq聚合酶或本领域内已知的其它DNA聚合酶。
重组所使用的方法取决于应该产生回交的同源性的基本程度。如果等同区域较大，可将Taq聚合酶与45-65℃的退火温度一起使用。如果等同区域较小，可将Klenow聚合酶与20-30℃的退火温度一起使用。本领域技术人员可以改变退火的温度以增加回交的数目。
可以在退火之前，与退火同时或退火之后将聚合酶加到随机核酸片段上。
在聚合酶存在下变性，变性恢复和保温的循环是指核酸的杂合或重组。将该循环重复预定的时间。优选的将该循环重复2-50次，更优选的重复10-40次。
得到的核酸是约50bp--约100kb的较大的双链多核苷酸，优选的较大多核苷酸是500bp--50kb。
该较大的多核苷酸片段可以含有互相串联的多个拷贝的核酸片段，该核酸片段的大小相同于模板多核苷酸。然后将该多联体片段消化为多拷贝的模板多核苷酸。结果产生大小几乎相同于模板多核苷酸的核酸片段群体。该群体是一个混合群体，其中在杂合之前已经向模板多核苷酸加入了具有等同区和异源区的单链或双链核酸片段。
然后将这些片段克隆到合适的载体中并将该连接混合物用于转化细菌。在克隆之前利用各种各样的方法包括PCR(美国专利№4683195和4683202)扩增单链合适片段而不是通过消化多联体可以从较大的多联体核酸片段获得单链核酸片段。
用于克隆的载体不是关键的，条件是它可以接受所需大小的DNA片段。如果需要该DNA片段进行表达，该克隆载体应该进一步含有与该DNA片段插入位点相连接的转录和转译信号以使该DNA片段在宿主细胞中表达。优选的载体包括pUC质粒系列和pBR质粒系列。
得到的细菌群体包括含有具有随机突变的许多重组DNA片段。对该混合群体进行检测以鉴别所需的重组核酸片段。选择方法取决于所需DNA片段。
例如如果需要为具有与配体的结合效力增强的蛋白质编码的DNA片段，采用本领域内已知的方法(即panning，亲和层析)检测由该群体或文库中每个DNA片段表达的蛋白质结合到配体上的能力。如果需要编码抗药性增加蛋白质的DNA片段，检测由该群体或文库中每个DNA片段表达的蛋白质授予宿主生物体抗药性的能力。本领域内技术人员借助于所需蛋白质的有关知识，很容易检测该群体以鉴别授予该蛋白质所需特性的DNA片段。
本发明涉及本领域内技术人员可以使用有关噬菌体展示系统，在该系统中在噬菌体的表面表达融合蛋白形式的蛋白质片段(Pharmacia，Milwaukee WI)。在导致融合蛋白转录的位点处将重组DNA分子克隆到噬菌体DNA中，该融合蛋白的一部分是由重组DNA分子编码的。含有重组核酸分子的噬菌体在细胞中进行复制或转录。该融合蛋白的前导序列指导该融合蛋白运输到噬菌体颗粒的顶端。因此由重组DNA分子部分编码的融合蛋白陈列于噬菌体颗粒上，采用上面描述的方法可以检测和选择。
本发明进一步涉及用来自第一群体的亚群体的核酸片段进行多个循环的核酸杂合，所说亚群体含有编码所需重组蛋白的DNA。按这种方式，可以获得有较高结合亲和力或酶促活性的蛋白质。
本发明还涉及用野生型核酸片段和来自第一或随后轮次的核酸杂交的核酸亚群体的混合物可以进行多个循环的核酸杂合，以便从亚群体中去除静止突变。
可以利用任何来源的纯化形式的核酸作为起始核酸。因此该方法可以利用DNA或RNA包括信使RNA，其中DNA或RNA可以是单链或双链。另外，可以利用含有各一条链的DNA-RNA杂合体。依据待诱变的核酸序列的大小，所说核酸序列可以是各种长度。优选的特定的核酸序列是50-50000bp。还涉及在本发明的方法中可使用含有编码所需蛋白质的核酸的完整载体。
可以从任何来源例如从质粒如pBR322，从克隆DNA或RNA或从来自细菌，酵母，病毒和高等生物如植物或动物的天然DNA或RNA获得核酸。可以从血液或组织材料中提取DNA或RNA。通过利用多核苷酸链反应(PCR)进行扩增可以获得模板多核苷酸(美国专利№4683202和4683195)。另一种方法，在存在于细胞中的载体中含有该核苷酸，并且通过培养该细胞和采用本领域内已知的方法从该细胞中提取核酸可以获得足够量的核酸。
采用本发明的方法可以利用任何特定的核酸序列生产突变群体。只需要在本发明方法之前存在或制造特定核酸序列的突变序列的小群体。
可采用许多不同的方法制备含有突变的特定核酸序列的起始小群体。采用易于产生错误的PCR可制备突变。易于产生错误的PCR利用低精确性的聚合条件以在整个序列长度内随机导入低水平的点突变。另一种方法，采用寡核苷酸定位诱变方法可将突变导入到模板多核苷酸中。在寡核苷酸定位诱变方法，利用限制性酶消化从该多核苷酸上去除短序列多核苷酸并且用一个合成多核苷酸取代，其中该合成的多核苷酸的各个碱基已经从原始序列发生变化。还可采用化学诱变方法改变该多核苷酸。化学诱变包括例如亚硫酸钠，亚硝酸羟胺，肼或甲酸。作为核酸前体类似物的其它试剂包括亚硝基胍，5-溴代尿嘧啶，2-氨基嘌呤或者吖啶。通常将这些试剂加到PCR反应中替代核酸前体从而使该序列突变。还可以使用插入试剂例如副核黄素，吖啶黄素，喹吖因等等。通过用X射线或紫外光照射也可以获得多核苷酸序列的随机诱变。通常，将如此诱变的质粒DNA或DNA片段导入到大肠杆菌中并且繁殖成突变质粒集合或文库。
另一种可用的方法找到了天然存在的特定核酸的混合小群体，它们由相同基因的不同等位基因或者不同相关种类的相同基因(即同族基因)组成。另一种可选择的方法，它们是一个种类存在的相关DNA序列，例如免疫球蛋白基因。
一旦产生了特定核酸分子的混合群体，可直接使用该多核苷酸或者利用本领域内熟知的技术将它插入到合适克隆载体中。
载体的选择依赖于多核苷酸序列的大小和本发明方法使用的宿主细胞。本发明的模板可以是质粒，噬菌体，粘粒，噬菌粒，病毒(例如反录病毒，副流感病毒，疱疹病毒，呼肠病毒，副粘病毒等等)或其选定的部分(例如包膜蛋白，针尖糖蛋白，衣壳蛋白)。例如，粘粒和噬菌粒是优选的，其中待诱变的特定核酸序列较大，因为这些载体能够稳定地繁殖大核酸片段。
如果将特定的核酸序列的混合群体克隆到一个载体中，通过将该各个载体插入到宿主细胞中并且使该宿主细胞扩增载体可以扩增该群体。这种方法称为克隆扩增，因为当核酸序列的绝对数增加时，突变数没有增加。
实用性
可以在未知序列的集合体中实施本发明的DNA杂合方法。通过在任何特定位点上将任何序列混合物加入到重组混合寡核苷酸(具有与被重组的序列同源的末端)，而将该序列混合物掺入到另一序列混合物中，因此本发明还涉及将合成寡核苷酸，PCR片段或者整个基因的混合物在限定的位置上混合到另一序列中。一个序列(混合物)的插入不依赖于一个序列插入到该模板的另一部分中。因此重组程度所需同源性，和文库的多样性可独立地和同时随重组DNA的长度而变化。
将两种基因混合的方法可用于使来自鼠杂交瘤的抗体具有人的特性。将两种基因混合或者将突变序列插入到基因中的方法可用于任何治疗上使用的蛋白质，例如白介素I，抗体，tPA，生长激素等等。该方法也用于任何核酸例如启动子或者内含子或者基因的3′未转移区或5′未转移区以提高表达或改变所表达蛋白质的特异性。该方法也可用于使核酶或aptamers。
杂合要求存在使多种多样的区域分离开的同源区。类似于支架状的蛋白质结构特别适合于杂合。当显示介导特定结合的相对非限制环时，保守的支架状蛋白确定了由自我联系进行的总体折迭。这样的支架状蛋白的例子是免疫球蛋白β管以及4个螺旋线束(24)。这种杂交方法结合各种组合的用于结合的突变序列可以制备类似支架状的蛋白。
体外杂合
用体外杂合也可以完成等量的一些标准遗传接合。例如，通过将突变体的核酸与野生型核酸重复混合，同时筛选所需的突变就可以完成’分子回交’。象在传统育种中一样，可以用这种方法将来自不同来源的表型结合到所选的背景中。例如，将其用于除去影响非选择特征(即免疫原性)的中性突变。因此，可以用来确定蛋白质中的哪个突变与生物活性的提高有关，而哪一个无关，用易出错误的诱变或盒诱变方法不能得到有利的突变。
可以从小随机片段混合物正确组装大的功能性基因。可以用所述反应对来自化石(25)的许多片段化的DNA进行基因组装。此外，可以将来自化石的随机核酸片段与来自相关种的相似基因的核酸片段结合。
还设想将本发明方法用于体外扩增进行各种研究和诊断应用所需的来自单个细胞的整个基因组。用PCR进行的DNA扩增在实践中限于约40kb的长度。用PCR扩增整个基因组，例如大肠杆菌(5,000kb)的，就需要约250个引物产生125个40kb片段。由于无法得到足够的序列资料，因而该方法不可行。另一方面，用DNAseI随机消化，然后用凝胶纯化小片段会得到许多可能的引物。用随机小片段的混合物在PCR反应中作为引物，单独或与整个基因组一起作为模板，会产生反向链反应，理论上结果产生含许多基因组拷贝的单个连环体。
当只使用随机片段时(见实施例2)可以得到拷贝数扩增100倍，平均片段大小大于50kb的结果。认为较大的连环体是由于许多较小片段的重叠而产生的。用合成引物得到的特异性PCR产物的性质无法与从未扩增的DNA得到的产物区别开。预期可以将所述方法用于基因组作图。
按照实践者的意愿，可以将杂合的多核苷酸制成随机的或非随机片段。
体内杂合
在体内杂合的实施方案中，在至少两种不同核酸序列存在于各宿主细胞的条件下，将特定核酸序列混合物导入到细菌或真核细胞中。可以用各种不同的方法，将所述片段导入到宿主细胞中。可以用本领域已知的方法，例如用氯化钙处理，用所述片段转化宿主细胞。如果所述片段被插入到噬菌体基因组中，则可以用含有特定核酸序列的重组噬菌体基因组转染宿主细胞。另外，可以用电穿孔，转染，脂染(lipofection)，biolistics，结合和类似的方法将所述核酸序列导入到宿主细胞中。
总之，在本实施方案中，所述特定的核酸序列将存在于能够在宿主细胞中稳定复制序列的载体中。另外，设想所述载体编码有关标记基因，以便可以筛选含有载体的宿主细胞。这样确保在导入到宿主细胞后，可以回收突变的特定核酸序列。然而，特定核酸序列的全部混合物不必均存在于载体序列上。相反，需要将足够数量的序列克隆到载体中以确保在所述片段导入到宿主细胞中后，各宿主细胞均含有一个携带至少一个存在于其上的特定核酸序列的载体。还设想不光是将特定核酸序列群体的子集克隆到载体中，该子集还已被稳定地整合到宿主细胞中。
已经发现，当将有相同区的两个片段插入到宿主细胞中时，在两个片段之间发生同源重组。在两个突变的特定核酸序列之间的所述重组在某些情况下，将会产生双或三突变体。
还发现如果某些突变的特定核酸序列存在于线性核酸分子上，则重组的频率会提高。因此，在优选的实施方案中，一些特定的核酸序列存在于线性核酸片段上。
转化后，将宿主转化体置于选择条件下，以鉴定含有带有所需性质的特定核酸序列的那些宿主细胞转化体。例如，如果需要对特定药物的增强抗性，则可以使转化宿主细胞经受增加浓度的特定药物，筛选那些产生突变蛋白质的转化体，所述蛋白质能够赋予增加的药物抗性。如果需要提高特定蛋白质与受体结合的能力，则从转化体诱导蛋白质的表达，然后用本领域熟知的方法，经配体结合试验检测所得的蛋白质以鉴定与配体结合能力提高的突变群体的子集。另外，可以在另一系统中表达所述蛋白质以确保适当的加工。
一旦鉴定了携带所需特征的第一重组的特定核酸序列的子集(女儿序列)，就可以使其进行第二个循环的重组。
在第二个重组循环中，可以将重组的特定核酸序列与源突变的特定核酸序列(亲本序列)混合，然后，重复上述的循环。用这种方法，可以鉴定一组第二重组的特定核酸序列，它们具有增强的特征或编码有增强的特性的蛋白质。可以按需要将所述循环重复数次。
还设想在第二或随后的重组循环中，可以进行回交。将所需的特定核酸序列与大量野生型序列混合，以便在转化后至少一个野生型核酸序列和突变的核酸序列存在于同一宿主细胞中，以完成分子回交。与野生型特定核酸序列的重组将会消除那些中性突变，所述中性突变会影响非选择特征如免疫原性，而不会影响所选择的特征。
在本发明的另一实施方案中，设想在第一轮中，在导入到宿主细胞之前，可以将特定核酸序列的子集制成片段。片段必须足够大以便含有与其他序列相同的一些区域，从而与所述其他序列进行同源重组。片段的大小为0.03kb到100kb，优选0.2kb到10kb。还设想在随后的数轮中，在导入到宿主细胞之前，将所有特定核酸序列而不是来自前一轮所选的序列裂解成片段。
可以用本领域已知的各种方法完成序列的片段化。可以在核酸序列的特定位点将所述序列制成随机片段或制成片段。通过断裂核酸或使其经严格的物理处理(例如剪切或辐射)或严格化学试剂(例如游离基团；金属离子；酸处理至脱嘌呤和裂解)可以得到随机片段。如果是DNA，也可以用DNase或类似的核酸酶得到随机片段。可以利用限制酶在特定的位点裂解而得到成片段的序列。制成片段的序列可以是单链或双链的。如果所述序列原本是单链的，那么在将其导入到宿主细胞之前可以用热、化学物质或酶将其变性。分开核酸链的合适的反应条件是本领域熟知的。
可以无限地重复本方法的步骤，但只受限于可得到的可能的突变体数目。在一定的循环数后，会得到所有可能的突变体，再循环就是多余的。
在一个实施方案中，重复重组相同的突变的模板核酸，然后筛选有所需特征的重组体。
因此，将突变模板核酸的起始池或群体克隆到能够在细菌，如大肠杆菌中复制的载体中。不需要特定的载体，只要它能够在大肠杆菌中能自我复制。在优选的实施方案中，设计所用的载体以便表达并生产由与载体相连的突变特定核酸编码的任何蛋白质。优选还有含有编码选择性标记的基因的载体。
将含有突变核酸序列池的载体群体导入到大肠杆菌宿主细胞中。可以经转化，转染或用噬菌体感染将载体核酸序列导入。用于转化细菌的载体浓度就是导入到各细胞中的载体量。一旦存在于细胞中后，同源重组的效率就是在不同载体之间发生的同源重组。这样会产生突变体(女儿)，它含有不同于源初亲本突变序列的突变的组合。
然后克隆复制宿主细胞并根据在载体上存在的标记基因进行筛选。只有含有质粒的那些细胞才能在选择条件下生长。
接着检测含载体的宿主细胞是否存在有利的突变。例如，如果待选的基因是一种改进的药物抗性基因，所述检测可由在选择的压力下涂布细胞组成。如果载体表达由突变的核酸序列编码的蛋白质，那么所述选择可包括表达所编码的蛋白质，分离所述蛋白质并检测所述蛋白质以确定例如它是否以提高的效率与所需的配体结合。
一旦鉴定了赋予所需特征的特定女儿突变核酸序列，就分离已与载体相连的核酸或从所述载体中分离核酸。然后将所述核酸与第一或亲本群体核酸混合，重复所述循环。
已经表明利用本方法可以筛选具有提高的所需特性的核酸序列。
在另一实施方案中，将第一代突变体保持在细胞中，然后将亲本的突变序列再加到所述细胞中。因此，按上述完成了实施方案I的第一循环。但是，在鉴定女儿核酸序列后，保留含有这些序列的宿主细胞。
将亲本突变的特定核酸序列(不论作为片段还是被克隆到相同的载体中)导入已含有女儿核酸的宿主细胞中。使得在细胞中发生重组，用上述方法筛选下一代重组体或孙女代。
可以将该循环重复许多次直到得到具有所需特征的核酸或肽。设想在随后的循环中，被加到优选突变体中的突变序列群体来自亲本突变体或任何后代。
在另一实施方案中，本发明提供了进行所得重组体的特定核酸的“分子”回交的方法，以便消除任何中性突变。中性突变是未赋予核酸或肽以所需特征的突变。但是，所述突变常常赋予核酸或肽以不需要的特征。因此，需要消除这样的中性突变。本发明的方法就提供了完成此项工作的方法。
在本实施方案中，用本实施方案的方法得到有所需特征的突变核酸后，分离所述核酸，含有所述核酸的载体或含所述载体的细胞。
然后将所述核酸或载体导入带有大量野生型核酸的宿主细胞中。使得突变体的核酸和野生型的核酸序列进行重组。将所得的重组体置于与突变体核酸相同的选择条件下，只有保留了所需特征的那些重组体才会被选择。不提供所需特征的任何不活动突变型通过与野生型DNA重组而被丢失。可以重复所述循环直到消除了所有不活动突变型。
因此可在分子回交中使用本发明的方法，以便消除不必的或不活动的突变。
实用性
可以对未知突变体的池或特定核酸片段或序列的等位基因盲目地使用本发明的体内重组方法。然而不必知道特定核酸片段的实际DNA或RNA序列。
在一个混合的基因群体内使用重组的方法可用于产生任何有用的蛋白质，例如白细胞介素I，抗体，tPA，生长激素等。可以用所述方法生产具有改变的特异性或活性的蛋白质。还可以用所述方法生产突变核酸序列，例如，启动子区，内含子，外显子，增强子序列，基因的3’非翻译区或5’非翻译区。因此可以用该方法产生高表达率的基因。还可用该方法研究重复DNA序列。最后，可以用该方法突变核酶或aptamers。
在蛋白质中分开多样性区的支架状(scaffold)样区特别适用于本发明.1的方法。保守的支架状区通过自我相连而确定所有的折叠，同时显示了介导特异性结合的相对不受限制的环。所述支架状区的实例是免疫球蛋白β管(barrel)和四螺旋束(bundle)。可以用本发明的方法生产具有用于结合的突变序列的不同组合的支架状区样蛋白质。
用本发明方法还可以完成一些标准遗传交配方法。例如通过将突变体核酸与野生型核酸重复混合，同时筛选所需的突变就可以完成“分子”回交。象在传统育种中一样，可以用本方法将不同来源的表型结合到所选的背景中。这对于例如除去影响非选择特征(即免疫原性)的中性突变是有用的。因此，可用其确定在蛋白质中哪个突变与增强的生物学活性有关，哪个无关。
肽展示方法
可以用本发明方法通过公开的体外和/或体内重组方法，并以任何组合杂合用肽展示方法筛选的多核苷酸序列，其中，有关的多核苷酸编码用于表型(如与预定受体(配体)的亲和性)筛选的展示的肽。
生物药物的研究和分子生物学的不断增长的重要性在于鉴定肽结构，包括肽的一级氨基酸序列，或于生物大分子相互作用的肽模拟。鉴定具有所需结构或功能特性，例如与预定生物大分子(如受体)的结合，包括筛选具有由所述肽氨基酸序列赋予的所需结构或功能特性的大文库或个体文库成员的肽。
除用直接的化学合成方法来产生肽文库外，还报道了数种重组DNA方法。一种是在噬菌体颗粒或细胞表面展示肽序列，抗体，或其他蛋白质。通常在这些方法中，各噬菌体颗粒或细胞是作为展示除天然噬菌体或细胞蛋白质序列之外的一种展示肽的个体文库成员。各噬菌体或细胞含有编码特定的展示肽序列的核苷酸序列信息；因此，通过确定所分离文库成员的核苷酸序列可以确定展示肽的序列。
熟知的肽显示方法包括在丝状噬菌体的表面上展示肽序列，通常是与噬菌体被膜蛋白融合。可将噬菌体文库与固定的，预先确定的大分子或小分子(如受体)一起保温，以便将呈现与固定大分子结合的肽序列的噬菌体颗粒与不呈现与预定大分子结合的肽序列的颗粒区别开。然后回收并复制与固定大分子结合的噬菌体颗粒(即文库成员)以扩增所选的噬菌体亚群体以便进行随后一轮的亲和富集及噬菌体复制。亲和富集及噬菌体复制数轮后，分离由此所选的噬菌体文库的成员，然后确定编码展示肽序列的核苷酸序列，从而鉴定与预定大分子(如受体)结合的肽的序列。所述方法在PCT专利公开号91/17271，91/18980，及91/19818和93/08278文本中有进一步的描述。
后一份PCT专利公开文本描述了用于展示肽配体的重组DNA方法，包括用各融合蛋白生产融合蛋白文库，所述融合蛋白由用于与预定大分子进行潜在结合的第一多肽部分，通常含有可变序列，以及结合DNA的第二多肽部分，例如编码单个融合蛋白的DNA载体组成。当在允许表达融合蛋白的条件下培养转化的宿主细胞时，融合蛋白就与编码它的DNA载体结合。将宿主细胞溶解后，用与在基于噬菌体的展示系统中筛选噬菌体颗粒极为相似的方法，对预定的大分子筛选融合蛋白/载体DNA复合物，以DNA载体在所选融合蛋白/载体DNA复合物中的复制和测序作为鉴定所选文库肽序列的基础。
生产肽和类似聚合物文库的其他系统有重组和体外化学合成方法双重功能。在这些杂交方法中，用无细胞的酶促机制完成文库成员(即肽或多核苷酸)的体外合成。在其中的一种方法中，利用改变选择和PCR扩增的轮次来选择具有与预定蛋白质或预定染料分子结合的能力的RNA分子(Tuerk and Gold(1990)Science249：505；Ellington and Szostak(1990)Nature 346：818)。相似的技术用于鉴定与预定的人转录因子结合的DNA序列(Thiesen and Bach(1990)Nucleic Acids Res.18：3203；Beaudry and Joyce(1992)Science 257；635；PCT专利92/05258和92/14843公开文本)。以相似的方式，已经用体外翻译技术合成所需的蛋白质并且作为生产大肽文库的方法。通常依赖于体外翻译，一般包括稳定的多核糖体复合物的这些方法在PCT专利公开88/08453，90/05785，90/07003，91/02076，91/05058和92/02536文本中有进一步的描述。申请人描述了其中文库成员含有融合蛋白的方法，所述融合蛋白含有具有DNA结合活性的第一多肽部分和具文库成员独特肽序列的第二多肽部分。所述方法适用于无细胞体外选择方式。
展示的肽序列其长度可以有所不同，通常为3-5000个氨基酸长或更长，一般为5-100个氨基酸长，且经常为8-15个氨基酸长。文库可以含有具有不同长度的展示肽序列的文库成员，或可含有固定长度的展示肽序列的文库成员。部分或所有展示的肽序列可以是随机的，伪随机的，限定的核心的，固定的等。本发明的展示方法包括用于体外和体内展示单链抗体的方法，例如，在多核糖体上的新单链Fv或在噬菌体上展示的单链Fv，所述方法可以大规模筛选含有较大可变区序列和结合特异性多样性的单链Fv文库。
本发明还提供随机，伪随机和限定的序列骨架肽文库以及生产并筛选这些文库以鉴定有用化合物(如肽，包括单链抗体)的方法，所述化合物以所需模式与受体分子或所需抗原决定簇或修饰肽的基因产物或的RNA结合。从肽文库成员的文库生产随机，伪随机和限定的序列骨架肽，包括肽文库成员附着于展示肽所来自的多核苷酸模板的展示肽或展示的单链抗体。附着模式随本发明的具体实施方案而不同，而且可包括包被在噬菌体颗粒中或掺入到细胞中。
利用亲和富集的方法可以筛选很大的肽和单链抗体文库，并筛选编码所需肽或单链抗体的多核苷酸序列。然后分离多核苷酸序列并杂合以便将所选肽(或其预定部分)或单链抗体(或仅其VH，VL或CDR)的氨基酸序列组合性重组。用这些方法，人们可以鉴定的由于具有对分子的所需的结合亲和性，肽或单链抗体并开发杂合方法以迅速地集中于所需的高亲和肽或scFv(单链Fv)。然后用适用于任何用途(例如治疗或诊断)的常规方法大量合成肽或抗体。
本发明的显著优点是事先不需要考虑预期的配体结构以用来分离所需的肽配体或抗体。所鉴定的肽会有生物学活性，这意味着包括对所选受体分子的特异性结合亲和性，在某些情况下，还包括阻断其他化合物结合、刺激或抑制代谢途径、作为信号或信使、刺激或抑制细胞活性等的能力。
本发明还提供杂合通过亲和筛选展示有新肽(包括单链抗体)的多核糖体的文库而选择的多核苷酸池的方法，所述新肽为结合预定受体(如哺乳动物蛋白类受体，例如peptidergic激素受体，细胞表面受体，与其他蛋白质结合以形成细胞内蛋白质复合物，如异二聚物等的细胞内蛋白质等)或抗原决定簇(例如固定的蛋白质，糖蛋白，寡糖等)的文库成员。
收集用这些方法中的任何方法在第一轮筛选(通常用对受体，如配体结合的亲和选择)中筛选的多核苷酸序列，然后在体外和/或体内重组杂合所述池以生产含有重组的所选多核苷酸序列的群体。使重组的所选多核苷酸序列经至少一次下一轮筛选。在后几轮中筛选的多核苷酸序列可被直接地使用，测序，和/或经一或多轮的杂合和选择。也可以将所选的序列与编码中性序列(即对结合无实质性影响)的多核苷酸序列回交，例如通过与野生型或实质上等同于所选序列的天然产生的序列回交来生产类似天然的，免疫原性较低的功能肽。总之，在回交过程中，用随后的选择保留与预定受体(配体)的结合特性。
在杂合所选的序列之前或同时，可以诱变所述序列。在一个实施方案中，将所选的文库成员克隆在原核载体(如质粒，噬菌粒，或噬菌体)中，其中产生代表不同文库成员的个体菌落(或噬菌斑)的集合。然后对单个的所选的文库成员进行操作(例如经定点诱变，盒诱变，化学诱变，PCR诱变等)以产生代表序列多样性的核心文库成员的集合，所述序列多样性以所选的文库成员为基础。可以对单个所选的文库成员或池的序列进行操作以掺入或者在所述单个所选文库成员序列的片段上，或在其全长上掺入。随机突变，伪随机突变，限定的核心突变(即含有变异体和非变异体残基位置和/或含有可含选自限定氨基酸残基子集的变异体残基位置)，密码子为基础的突变等，然后用本文所述的体外和/或他内重组杂合来杂合诱变后的所选文库成员。
本发明还提供含有本发明单个文库成员多样性的肽文库，其中(1)所述多样性的各文库成员含有通过杂合所选序列的池而产生的序列，和(2)各单个文库成员含有在所述多样性中不同于其他单个文库成员的可变肽片段序列或单链抗体序列的可变肽片段序列或单链抗体序列(尽管由于不均匀扩增，随机可能性等，某些文库成员可能以多于1个拷贝/文库存在)。
本发明还提供产物的生产方法，其中具有(或编码的肽具有)预定结合特异性的所选的多核苷酸序列经下列过程形成：(1)筛选抗预定受体(如配体)或抗原决定簇(如抗原大分子)的展示肽或展示的单链抗体文库，然后鉴定和/或富集与所述预定受体或抗原决定簇结合的文库成员以产生所选文库成员的池，(2)通过重组与预定的抗原决定簇结合所选的文库成员(或扩增或克隆其拷贝)并且由此从文库中分离和/或富集而进行杂合以产生杂合的文库，和(3)筛选抗预定受体(如配体)或抗原决定簇(如抗原大分子)的杂合的文库并鉴定和/或富集与预定受体或抗原决定簇结合的文库成员以产生所选杂合文库成员的池。
抗体展示和筛选方法
可以用本发明方法通过任何公开的体外和/或体内重组方法并以任何组合杂合用抗体展示方法筛选的多核苷酸序列，其中有关的多核苷酸编码用于表型(例如用于结合预定抗原(配体)的亲和性)筛选的展示的抗体。
已经设计了各种分子遗传学方法以得到由极大量可存在于免疫球蛋白链上的不同可变区所代表的大量免疫学组分。天然产生的种系免疫球蛋白重链基因座位由位于不同(D)片段基因串联排列上游的可变(V)片段基因的分开的串联排列组成，D片段基因自身位于连接(J)区基因串联排列的上游，J区位于恒定(C)区基因的上游。在B淋巴细胞发育的过程中，发生V-D-J重排，其中通过重排形成融合的D-J片段，然后又与V片段重排形成V-D-J相连的产物基因，从而形成重链可变区基因(VH)，如果是生产性重排，V-D-J编码重链的功能可变区(VH)。相似地，轻链基因座位上，数个V片段之一与数个J片段之一重排以形成编码轻链可变区的基因。
免疫球蛋白上许多可能的可变区的所有组成成分部分来自在B细胞发育中重排过程中V和J片段(在重链情况下，和D片段)连接的大量组合性可能性。在重链可变区中其他的序列多样性产生于V-D-J连接过程中非均一地重排和N区的增加。此外，特异性B细胞克隆的抗原选择作用选择在重链和轻链可变区之一或两者中有非种系突变的高亲和突变体；这是一种称为“亲和性成熟”或“亲和性加强”的现象。通常，这些“亲和性加强”集结在可变区的特定区域，常常在互补性决定区(CDRs)。
为了克服通过抗原-刺激的B细胞发育(即免疫)生产和鉴定高亲和性免疫球蛋白的许多限制，已经研究了各种可操作的原核表达系统以生产可用于筛选高亲和性的抗特异性抗原的抗体的组成型抗体文库。在大肠杆菌和噬菌体系统中表达抗体的最近进展(见“Altermative Peptide Display Methods”同下文)已经产生了实际上可通过从表征的杂交瘤克隆抗体基因或用抗体基因文库(如从Ig cDNA)重新筛选得到任何特异性的可能性。
在可筛选为噬菌斑或溶原菌落的噬菌体λ表达系统中已经生产了抗体的组成型文库(Huse et al.(1989)Science 246：1275；Caton and Koprowski(1990)Proc.Natl.Acad.Sci.(U.S.A.)87：6450；Mullinax et al(1990)Proc.Natl.Acad.Sci.(U.S.A.)87：8095；Persson et al.(1991)Proc.Natl.Acad.Sci.(U.SA.)88：2432)。已经描述了噬菌体抗体展示文库和λ噬菌体表达文库的各种实施方案(Kang et al.(1991)Proc.Natl.Acad.Sci.(U.S.A.)88：4363；Clackson et al.(1991)Nature 352：624；McCafferty et al.(1990)Nature 348：552；Burton et al.(1991)Proc.Natl.Acad.Sci.(U.S.A.)88：10134；Hoogenboom et al.(1991)Nucleic Acids Res.19：4133；Chang et al.(1991)J.Immunol.147：3610；Breitling et al(1991)Gene 104：147；Marks et al.(1991)J.Mol.Biol.222：581；Barbas et al.(1992)Proc.Natl.Acad.Sci.(U.S.A.)89：4457；Hawkins and Winter(1 992)J.Immunol.22：867；Marks et al.(1 992)Biotechnology 10：779；Marks et al.(1992)J.Biol.Chem.267：16007；Lowman et al.(1991)Biochemistry 30：10832；Lerner et al.(1992)Science 258：1313；均引入本文作为参考)。通常，用固定(如通过与层析树脂共价结合以便用亲和层析富集反应性噬菌体)和/或标记(如以筛选噬菌斑或挑取菌落)的受体(如多肽，碳水化合物，糖蛋白，核酸)筛选噬菌体抗体展示文库。
一种特别有利的方法是利用所谓单链片段可变(scFv)文库(Marks et al.(1992)Biotechnology 10：779；Winter G and Milstein C(1991)Nature 349：293；Clackson et al.(1991)，在所引的书中；Marks et al.(1991)J.Mol.Biol.222：581；Chaudhary et al.(1990)Proc.Natl.Acad.Sci.(U.SA.)87：1066；Chiswell et al.(1992)TIBTECH 10：80；McCafferty et al.(1990)在所引的书中；and Huston et al.(1988)Proc.Natl.Acad.Sci.(U.S.A.)85：5879)。已经描述了各种展示在噬菌体被膜蛋白上的scFv文库的实施方案。
从1988年开始，用抗体基因工程方法就很容易生产Fv片段的单链类似物和其融合蛋白。通常第一步包括得到编码具有所需结合特性的VH和VL区的基因；这些V基因可以从特定的杂交瘤细胞系中分离，或从组成型V-基因文库中筛选或通过V基因合成来制备。通过将成分V基因和编码适当设计的接头肽，例如(Gly-Gly-Gly-Gly-Ser)3或接头肽等同物的寡核苷酸连接起来形成单链Fv。接头桥接第一个V区的C-末端和第二个的N-末端，连接成VH-接头-VL或VL-接头-VH。理论上，scFv结合位点可以精确地复制其亲本抗体结合位点的亲和性和特异性。
因此，scFv片段由通过灵活的接头肽连接到一单链多肽中的VH和VL区组成。组装成scFv基因后，将其克隆到噬菌粒中，然后在M13噬菌体(或相似丝状噬菌体)的顶部表达为与噬菌体pIII(基因3)被膜蛋白融合的融合蛋白。通过培养(panning)重组噬菌体来富集表达所需抗体的噬菌体，所述重组噬菌体带有与预定抗原决定簇(例如靶抗原，受体)结合的scFv群体。
连接的文库成员的多核苷酸提供了在筛选或选择方法后复制文库成员的基础，同时还提供了通过核苷酸测序确定展示的肽序列或VH和VL氨基酸序列相同性的基础。抗原在适宜的表达系统中克隆并表达展示的肽或单链抗体(如scFv)和/或其VH和VL区或其CDRs。经常将编码分离的CDRs的VH和VL区与编码恒定区(CH和CL)的多核苷酸相连以形成编码完整抗体，(如嵌合的或完全的人)抗体片段等。经常将编码分离的CDRs的多核苷酸移植到编码适宜可变区骨架(和可选择的恒定区)多核苷酸中以形成编码完整抗体，(如嵌合的或完全的人的)抗体片段等。可以用抗体通过免疫亲和层析分离制备量的抗原。所述抗体的其他用途是诊断和/或确定疾病的阶段(如瘤形成)，用于治疗疾病，例如瘤形成，自体免疫疾病，AIDS，心血管病，感染等。
已经报告了各种方法以增加scFv文库组成型的多样性，从而扩大结合种类(个体基因型谱)的所有组成。用PCR可以迅速从特定杂交瘤或作为基因文库从非固定的细胞中克隆可变区，在可以组合的VH和VL盒的分类中提供组成型多样性。此外，例如通过随机，伪随机或定位诱变可将VH和VL盒自身多样化。通常，在或靠近互补性决定区(CDRs)，经常在第三个CDR，CDR3将VH和VL盒多样化。由于有易差错的PCR和化学诱变(Deng et al.(1994)J.Biol.Chem.269：9533)，已经表明酶促反向PCR诱变是一种简单可行的构建相对大的scFv定点诱变突变体文库的方法(Stemmer et al.(1993)Biotechniques 14：256)。Riechmann等人((1993)Biochemistry 32：8848)使用通过简并寡核苷酸PCR和所得scFv突变体的噬菌体展示的定点随机化列出了半推理设计的抗体scFv片段。Barbas等人((1992)，在所引的书中)试图回避限制所有组分大小的问题，而该问题是由于使用偏移的可变区序列通过随机化在人破伤风类毒素-结合Fab的合成CDR区中的序列而造成的。
CDR随机化有可能仅对重链CDR3的约1×1020就产生CDRs，以及大约相似量的重链CDR1和CDR2突变体和轻链CDR1-3突变体。单独或一起考虑，重链和/或轻链CDR随机化的组成要产生抑制量的噬菌体克隆以生产单独代表所有可能组合的一个克隆文库，其中大多数是非结合的。用目前的转化技术和噬菌体展示系统要生产这样大量的一级转化体并不容易。例如Barbas等人(1992，在所引的书中)只产生了5×107个转化体，这只代表了所有随机化CDRs文库潜在多样性的很小的部分。
尽管有这些实质性的限制，但scFv的噬菌体展示还是产生了各种有用的抗体和抗体融合蛋白。已经表明双特异性单链抗体介导有效的肿瘤细胞溶解(Gruber et al；(1994)J.Immunol.152：5368)。已表明抗-Rev scFv的细胞内表达抑制HIV-1的体外复制(Duan et al.(1994)，Proc.Natl.Acad.Sci.(USA)91：5075)，抗-p21rasscFv的细胞内表达抑制Xenopus卵母细胞的减数分裂成熟(Biocca et al.(1993)Biochem.Biophys.Res.Commun.197：422)。还报告了用于诊断HIV感染的重组scFv，表明了scFv的诊断实用性(Lilley et al.(1994)J.Immunol.Meth.171：211)。也已经报告了scFv与第二多肽，如毒素或血纤维蛋白激活蛋白相连的融合蛋白(Holvost et al.(1992)Eur.J.Biochem.210：945；Nicholls et al.(1993)J.Biol.chem.268：5302)。
如果可能生产具有更多抗体多样性并克服常规CDR诱变和随机化方法的许多限制的scFv文库，就可以使适用于治疗和诊断用途的scFv抗体的数量和质量得到很大的提高，所述的常规CDR诱变和随机化方法只能覆盖潜在序列组合中很小的部分。为了达到这一目的，用本发明的体外和体内杂合方法重组已经从核酸得到(通常经PCR扩增或克隆)的CDRs，所述核酸是从筛选的展示抗体得到的。可以在细胞上，噬菌体颗粒上，多核糖体或其中有抗体与编码它的核酸相连的适宜的抗体展示系统上展示所述的展示抗体。在改变中，CDRs开始是从生产抗体的细胞(例如来自免疫的野生型小鼠，人或能够生产WO92/03918，WO93/12227和WO94/25585中的人抗体的转基因小鼠的浆细胞/脾细胞)，包括从其得到的杂交瘤的mRNA(或cDNA)得到的。
收集用任何这些方法在第一轮筛选(通常通过亲和性筛选与抗原(例如，配体)结合的展示的抗体)中筛选的多核苷酸序列，然后经体外和/或体内重组进行杂合，特别是杂合CDRs(通常杂合重链CDRs和其他重链CDRs以及轻链CDRs和其他轻链CDRs)以得到杂合的池，所述池含有重组的筛选过的多核苷酸序列群体。将重组的选择的多核苷酸序列在选择方式中表达随后再经至少一轮筛选。直接使用，测序在随后的选择中筛选的多核苷酸序列，和/或使其经一次或多次的杂合和筛选过程，直到得到具有所需结合亲和性的抗体。也可以将所选的序列与编码中性抗骨架序列的多核苷酸序列(即对结合抗原没有实质性的功能影响)回交，例如通过与人可变区骨架回交以得到类似人的抗体序列。总而言之，在回交过程中，用随后的选择作用来保留与预定抗原结合的特性。
另外，或与注明的改变相结合，可以改变靶抗原决定簇的效价以控制所选scFv文库成员的平均结合亲和性。可以将靶抗原决定簇以不同的密度，通过例如包括竞争抗原决定簇，通过稀释或通过本领域已知的其他方法，与表面或底物结合。可以用高密度(效价)的预定抗原决定簇富集具有相对低亲和性的scFv文库成员，而低密度(效价)可优选地富集较高亲和性的scFv文库成员。
为了产生不同的可变片段，通过连接到预定位点(如CDR)插入编码随机，伪随机，或限定的核心序列组肽序列的合成寡核苷酸插入集合。相似地，用定点诱变，CDR-取代等通过突变CDR来扩展单链抗体盒的一个或多个CDRs的序列多样性。在杂合前，可以在克隆和扩增的宿主中增殖所得的DNA分子，或将其直接使用(即可以避免在宿主细胞中增殖时可能发生的丢失)，然后杂合所选的文库成员。
通过亲和富集技术从所述文库中筛选编码所需可变片段肽序列或所需单链抗体的肽/多核苷酸复合物(文库成员)。利用固定的大分子或对所需肽序列特异性的抗原决定簇，例如受体，其他大分子，或其他抗原决定簇达到所述目的。重复亲和筛选方法可以富集编码所需序列的文库成员，然后将其分离以进行收集和杂合，测序和/或进行进一步增殖和亲和富集。
通过洗涤除去没有所需特异性的文库成员。确定对于所需的各肽序列或单链抗体及固定的预定大分子或抗原决定簇所需的洗涤程度和严谨度。通过调节结合温育和随后洗涤的条件，可以在一定程度上控制回收的新肽/DNA复合物的结合特征。在固定的大分子亲和性的特定范围内，将选择用于新肽/DNA复合物的温度，pH，离子强度，二价阳离子浓度以及洗涤体积和持续的时间。以慢解离速率为基础的选择(通常预测到高亲和性)是最常用的实施途径。通过在存在饱和量的游离预定大分子条件下继续保温或通过增加洗涤体积，次数和时间长度来达到所述目的。在各情况下，要预防解离的新肽/DNA或肽/RNA复合物的再结合，而且随着时间的增加，回收亲和性越来越高的新肽/DNA或肽/RNA复合物。
也可以使用对结合和洗涤过程有其他改进的方法以发现具有特殊特征的肽。某些肽的亲和性依赖于离子强度或阳离子浓度。当需要从肽中除去蛋白质的条件柔和时，这对于在各种蛋白质的亲和纯化中使用的肽来说是有用的特征。
一种改变是利用多种结合靶(多种抗原决定簇，多种受体)以便同时筛选scFv文库，筛选具有不同结合特异性的多重性的scFv。由于scFv文库的大小经常限制潜在scFv序列的多样性，因此，一般我们希望scFv文库尽可能地大。出于时间和经济上的考虑，产生许多很大的多核糖体scFv-展示文库会受到限制。为了避免这个实际问题，在单个文库中可以同时筛选多种预定的抗原决定簇(受体)，或对大量抗原决定簇进行依次筛选。在一种改变中，可以混合在不同珠(或珠组)上编码的多种靶抗原决定簇，然后在适宜的结合条件下与多核糖体-展示scFv文库保温。然后通过亲和筛选，收集的含有多种抗原决定簇的珠来分离scFv文库成员。通常，随后的亲和筛选过程包括混合相同的小珠，其子集或含有仅一种或两种抗原决定簇的小珠。所述方法提供了有效的筛选，而且与实验室自动化，批量加工，筛选方法的高处理量相协调。
在本发明中可以使用各种技术以便使肽文库或单链抗体文库多样化，或在杂合前或同时，使在较早的panning过程中发现的可变片段肽或VH，VL或CDRs周围多样化使其对预定大分子或抗原决定簇具有足够的结合活性。在一种方法中，将筛选的阳性肽/多核苷酸复合物(在较早的亲和富集过程中鉴定的那些)测序以确定活性肽的一致性。然后在这些活性肽的基础上合成寡核苷酸，在各步骤中通过低水平的所有碱基掺入以得到有轻微改变的一级寡核苷酸序列。将所述(轻微)简并寡核苷酸的混合物在适当的位置克隆到可变片段序列中。这种方法对起始肽序列产生了系统的控制的改变，然后可用于杂合。然而，它要求在诱变前对单个阳性新肽/多核苷酸复合物测序，因此对于扩大少量所回收的复合物的多样性并筛选具有更高结合亲和性和/或更高结合特异性的变异体来说是有用的。在一种改变中，经诱变PCR扩增所选的阳性肽/多核苷酸复合物(特别是可变区序列，在体外和/或体内杂合的扩增产物)并在测序前再进行一轮或多轮的筛选。对于单链抗体可以使用大致相同的方法以便通过在杂合前或同时将CDRs或相邻的骨架区多样化而扩大多样性并提高结合亲和性/特异性。如果需要，用能够与包括所选文库成员进行体外重组的诱变寡核苷酸抑制杂合反应。因此，可以将合成的寡核苷酸和PCR片段(用易于差错或高精确度方法合成的)混合物加到体外杂合混合物中，然后掺入到所得的杂合文库成员(杂合体)中。
本发明的杂合能够产生大的CDR-变异体单链抗体文库。产生所述抗体的一种方法是在杂合前或同时将合成的CDRs插入到单链抗体和/或随机化的CDR中。通过参考已知的人CDR的序列资料来筛选合成的CDR盒序列并且根据下列原则由实施慎重选择：合成的CDRs与已知CDR序列至少有40％的序列一致性，而且优选的与已知序列至少有50-70％的一致性。例如，在Kabat等人(1991，在所引的书中)所列的天然产生的人CDR序列的基础上，通过合成一批寡核苷酸序列来产生一批合成的CDR序列；计算合成CDR序列的池以使其编码与至少一种已知的天然产生的CDR序列有至少40％一致性的CDR肽序列。另外，可以比较一批天然产生的CDR序列以得到一致序列以便将在频率高的残基位置(即占已知CDR序列至少5％)所用的氨基酸掺入到合成CDRs的相应位置。通常比较并观察一些(如3到约50个)一致的CDR序列，将一致CDRs之间的天然序列变异列成表，然后合成一批编码CDR肽序列的寡核苷酸，所述CDR肽序列包括所有或大多数观察到的天然序列变异的置换。例如并不是为了限制，如果一批人VHCDR序列含有以Tyr，Val，Phe，或Asp为C-末端氨基酸，那么就可以设计合成CDR寡核苷酸序列的池以使所述C-末端CDR残基是这些氨基酸中的任何一种。在一些实施方案中，掺入除在CDR序列中的残基位置上的天然残基之外的残基：经常插入保守的氨基酸取代物，而且可以改变高达5个残基位置以掺入与天然CDR序列相比是非保守的氨基酸取代。可以将所述CDR序列用于一级文库成员(在第一轮筛选前)和/或可用其抑制所选文库成员序列的体外杂合反应。本领域专业人员很容易构建限定的和/或简并序列的所述池。
所述合成的CDR序列含有至少一个不是已知天然CDR序列的成员。实施者可任选包括或不包括相对于重链CDR N区的随机或伪随机序列部分；在V-D和D-J的连接中N区序列的范围是1个核苷酸至约4个核苷酸。一批合成的重链CDR序列含有至少约100个特有的CDR序列，通常至少为1,000个特有的CDR序列，优选地为至少约10,000个特有的CDR序列，经常多于50,000个特有的CDR序列；但是在一批中通常不超过1×106个特有的CDR序列，尽管有时存在1×107-1×108个特有CDR序列，特别是如果在天然人CDRs中，在没有保守氨基酸取代物的位置或在该位置保守氨基酸取代物稀少(即少于0.1％)的情况下，允许保守氨基酸取代。总之，在文库中所包括的特有CDR序列的数量不能超过所述文库中一级转化体预期数量的10倍以上。所述单链抗体通常可以以约至少1×107M-1的亲和性，优选有约至少5×107M-1的亲和性，更优选的约至少1×108M-1到约至少1×109M-1的亲和性或更高，有时高达1×1010M-1或更高的亲和性与预定抗原(如免疫原)结合。通常预定抗原是人蛋白质，例如人细胞表面抗原(如CD4，CD8，IL-2受体，EGF受体，PDGF受体)，其他人生物大分子(如血栓调节蛋白，蛋白质C，碳水化合物抗原，sialyl Lewis抗原，L-选择素(selectin))，或与人疾病无关的大分子(如细菌LPS，病毒颗粒衣壳蛋白或被膜糖蛋白)等。
抗原在各种系统中加工并表达具有所需特异性的高亲和单链抗体。例如已经在植物中生产了scFv(Firek et al.(1993)Plant Mol.Biol.23：861)，而且很容易在原核系统中制备(Owens RJ(1994)J.Immunol.Meth.168：149；Johnson S and Bird RE(1991)Methods Enzymol.203：88)。此外，可以用单链抗体作为构建完全抗体或其各种片段(Kettleborough et al.(1994)Eur.J.Immunol.24：952)。可以分离(如经PCR扩增或亚克隆)可变区编码序列并将其与编码所需人恒定区的序列相拼接以编码人抗体序列，所述抗体更适于免疫原性最小的的人治疗应用。在宿主细胞中表达含有所得的人编码序列的多核苷酸(例如在哺乳动物细胞中从表达载体开始)，然后将其纯化用于药物配方。
DNA表达构建体通常包括与编码序列可操作相连的表达控制DNA序列，包括天然相关的或异源的启动子区。优选的，表达控制序列是在能够转化或转染真核宿主细胞的载体中的真核启动子系统。一旦所述载体被插入到适宜的宿主中，就在适于高水平表达核苷酸序列的条件下保持所述宿主，然后收集纯化突变的“工程”抗体。
正如上文所述，在将DNA序列与表达控制序列可操作相连(即定位以确保结构基因的转录和翻译)后，在宿主中表达所述序列。这些载体通常作为附加体或宿主染色体DNA的整合部分在宿主生物体中是可复制的。总之，表达载体将含有选择性标记如，四环素或新霉素以便检测带有所需的DNA序列的那些转化细胞(见，如美国专利4,704,362，该文献引入本文作为参考)。
除象酵母这样的转化微生物外，也可以用哺乳动物组织细胞生产本发明的多肽(见Winnacker，“From Genes to Clones，”VCH Publishers，N.Y.，N.Y.(1987)，该文献引入本文作为参考)。实际上真核细胞是优选的，原因在于在本领域中已经研究了许多能够分泌完整免疫球蛋白的适宜的宿主细胞系，而且包括CHO细胞系，各种COS细胞系，Hela细胞系，骨髓瘤细胞系等，但最好是转化的B-细胞或杂交瘤。用于这些细胞的表达载体包括表达控制序列，如复制源点，启动子，增强子(Queen et al.(1986)Immunol.Rev.89：49)，和必需的加工信息位点，如核糖体结合位点，RNA拼接位点，多腺苷酸化位点以及转录终止序列。优选的表达控制序列源于免疫球蛋白基因，细胞肥大病毒，SV40，腺病毒，牛乳头状瘤病毒等的的启动子。
通过将增强子序列插入到载体中来提高真核DNA的转录。增强子是10-300bp的提高启动子转录的顺式作用序列。当在转录单位5′或3′时，增强子可以有效地提高转录。如果在内含子内或在其自身编码序列内，它们也是有效的。通常使用的表达增强子包括SV40增强子，细胞肥大病毒增强子，多形瘤增强子和腺病毒增强子。来自哺乳动物系统的增强子序列也是常用的，例如小鼠免疫球蛋白重链增强子。
哺乳动物表达载体系统通常还包括可选择的标记基因。适宜标记基因的实例包括二氢叶酸还原酶(DHFR)，胸苷激酶基因(TK)或赋予药物抗性的原核基因。前两种标记倾向于使用在未将胸苷加到生长培养基中就不能生长的突变体细胞系。根据其在非补充培养基中生长的能力来鉴定转化的细胞。作为标记的有用的原核药物抗性基因的实例包括赋予G418，霉酚酸和潮霉素抗性的基因。
根据细胞宿主类型，用熟知的方法，可以将含所需的DNA片段的载体转移到宿主细胞中。例如常用于原核细胞的氯化钙转染，而磷酸钙处理。脂染或电穿孔用于其他细胞宿主。用于转化哺乳动物细胞的其他方法包括使用聚凝胺，原生质体融合，脂质体，电穿孔和微注射(见，Sambrook et al.同上文)。
一旦表达后，可以用本领域的标准方法，包括硫酸铵沉淀，分离柱层析，凝胶电泳等纯化本发明的抗体，个体突变的免疫球蛋白链，突变的抗体片段和其他免疫球蛋白多肽(见Scopes R.，Protein Purification，Springer-Verlag，N.Y.(1982))。纯化后，按照部分或均一性的要求，将多肽用于治疗或研究和完成检测方法，没有荧光染色等(见，Immunological Methods，Vols.I and II，Eds.Lefkovits and Pernis，Academic Press，New York，N.Y.(1979 and 1981))。
用本发明方法生产的抗体可用于诊断和治疗。用以说明而不是限制为目的，可以用它们治疗癌，自体免疫疾病或病毒感染。为了治疗癌，通常抗体将与常在癌细胞上表达的抗原，例如erbB-2，CEA，CD33以及许多其他抗原和本领域专业人员熟知的结合成员结合。
酵母两杂交筛选试验
可以用杂合重组对通过筛选两杂交筛选系统来鉴定结合预定多肽序列的文库成员而得到的所选文库成员池多样化。收集所选的文库成员，然后进行体外和/或体内重组。然后在酵母两杂交系统中筛选杂合的池以筛选结合所述预定多肽序列(如SH2区)或结合其他预定多肽序列(如来自另一蛋白质的SH2区)的文库成员。
鉴定结合预定多肽序列的多肽序列鉴定方法已经使用了所谓“两杂交”系统，其中预定多肽序列存在于融合蛋白中(Chien et al.(1991)Proc.Natl.Acad.Sci.(USA)88：9578)。所述方法通过重建转录活化因子(Fields S and Song O(1989)Nature 340：245)，酵母Ga14转录蛋白来鉴定蛋白质-蛋白质之间的相互作用。通常，该方法是以酵母Ga14蛋白质的特性为基础的，所述蛋白质由导致DNA结合和转录活化的可分开的区组成。构建编码两种杂交蛋白质的多核苷酸并将其导入到酵母宿主细胞中，所述两种杂交蛋白，一种由与已知蛋白质的多肽序列融合的酵母Ga14 DNA结合区组成，另一个由与第二蛋白质的多肽序列融合的Ga14活化区组成。两种融合蛋白之间的细胞间结合重建了Ga14 DNA结合区和Ga14活化区，这样导致与Ga14结合位点可操作相连的报告基因(如lacZ，HIS3)的转录活化。通常用两杂交方法鉴定与已知蛋白质相互作用的新多肽(Silver SC and Hunt SW(1993)Mol.Biol.Rep.17：155；Durfee et al.(1993)Genes Devel.7：555；Yang et al.(1992)Science 257：680；Luban et al.(1993)Cell 73：1067；Hardy et al.(1992)GenesDevel.6；801；Bartel et al.(1993)Biotechniques 14：920；and Vojtek et al.(1993)Cell74：205)。但是，已经用改进的两杂交方法鉴定影响其与第二已知蛋白质结合的已知蛋白质的突变体(Li B and Fields S(1993)FASEB J.7：957；Lalo et al.(1993)Proc.Natl.Acad.Sci.(USA)90：5524；Jackson et al.(1993)Mol.Cell.Biol.13：2899；andMadura et al.(1993)J.Biol.Chem.268：12046)。还已经用两杂交系统鉴定了两个已知蛋白质结构区(Bardwell et al.(1 993)med.Microbiol.8：1177；Chakraborty et al.(1992)J.Bio.Chem.267：17498；Staudinger et al.(1993)J.Biol.Chem.268：4608；andMilne GT and Weaver DT(1993)Genes Devel.7：1755)或引起单个蛋白质寡聚的区(Iwabuchi et al.(1993)Oncogene 8：1693；Bogerd et al.(1993)J.Virol.67：5030)之间的相互作用。还用改进的两杂交系统研究蛋白水解酶的体内或(Dasmahapatra etal.(1992)Proc.Natl.Acad Sci.(USA)89：4159)。另外，可以用大肠杆菌/BCCP相互作用筛选系统(Germino et al.(1993)Proc.Natl.Acad Sci.(USA)90：933；GuarenteL(1993)Proc.Natl.Acad Sci.(USA)90：1639)鉴定相互作用的蛋白质序列(即异二聚的或形成更高级异多聚体的蛋白质序列)。收集用两杂交系统选择的序列，将其杂合，然后导入到用于随后一或多轮筛选以鉴定与含预定结合序列结合的多肽序列的两杂交系统中。将由此鉴定的序列进行比较以鉴定一致序列或一致序列核心。
从上述公开内容可以理解，本发明有广泛的应用。因此提供下列实施例是为了说明而不是限制。
在下列实施例中，下列缩写有下列含义。如果在下文未说明，那么所述缩写含有本领域已知的含义。
ml＝毫升
μl＝微升
μM＝微摩尔
nM＝毫微摩尔
PBS＝磷酸缓冲的盐
ng＝毫微克
μg＝微克
IPTG＝异丙基硫代-β-D-半乳糖苷
bp＝碱基对
kb＝千碱基对
dNTP＝三磷酸脱氧核苷酸
PCR＝聚合酶链反应
X-gal＝5-溴-4-氯-3-吲哚基-β-D-半乳糖苷
DNAseI＝脱氧核糖核酸酶
PBS＝磷酸缓冲的盐
CDR＝互补性决定区
MIC＝最小抑制浓度
scFv＝抗体的单链Fv片段
总之，在各种文献中描述了标准的重组DNA技术，例如Sambrook et al.，1989，Molecular Cloning：A Laboratory Manual，Cold Spring Harbor Laboratry；Ausubel et al.1987，Current Protocols in Moleculat Biology，Vols 1 and 2和补充，and Berger andKimmel，Methods in Enzymology，Volume 152，Guide to Molecular ClongingTechniques(1987)，Academic Press，Inc.Sam Diego，Ca，以上文献均全部引入本文作为参考。按照制造商的说明使用限制酶和多核苷酸修饰酶。使用ABI混合物，在Applied Biosystems Inc.Model 394 DNA合成仪上合成寡核苷酸。如果需要，根据实践者的需要选择扩增预定DNA序列的PCR引物。
                          实施例
实施例1.LacZα基因重装配
1)底物制备
重组反应的底物是来自pUC18(图2)(28；Gene BankNo.XO2514)的野生型LacZα基因的dsDNA聚合酶链反应(“PCR”)产物。引物序列是5′AAAGCGTCGATTTTTGTGAT3′(SEQ ID NO：1)和5′ATGGGGTTCCGCGCACATTT3′(SEQ ID NO：2)。根据厂家说明书以Wizard PCR制品(Promega，Madison WI)从PCR产物中去掉游离的引物。发现游离引物的清除是重要的。
2)DNAseI消化：
用溶于100μl[50mM Tris-HCl pH7.4，1mM MgCl2]中的0.15单位的DNAseI(Sigma，St.Louis MO)在室温下消化大约5μg DNA底物10-20分钟。消化的DNA在2％低熔点琼脂糖凝胶上电泳。经过电泳到DE81离子交换纸(Whatman，Hillsborough OR)上从2％低熔点琼脂糖凝胶上纯化10-70碱基对(bp)的片段。用1M NaCl从纸上洗脱出DNA片段并用乙醇沉淀。
3)DNA重装配：
将纯化的片段以10-30ng/μl的浓度重悬于PCR混合物(各0.2mM dNTP，2.2mM MgCl2，50mM KCl，10mM Tris-HCl pH9.0，0.1％Triton X-100，0.3μl Taq DNA聚合酶，总体积50μl)。在此时不加引物。在MJ Research(Watertown MA)PTC-150热循环器中使用的重组程序是94℃ 60秒，30-45个循环的[94℃ 30秒，50-55℃ 30秒，72℃ 30秒]及72℃ 5分钟。小片段重组进较大序列的PCR后在重组25，30，35，40和45个循环后取反应样品(图2)。
由于100-200bp片段的重组可产生正确大小的单个PCR产物，10-50碱基片段典型地产生正确大小的一些产物以及不均一分子量的产物。大多数这种大小不均一性似乎是由于产物末端是单链序列，因为限制性酶消化后获得正确大小的单一带。
4)有引物的PCR
将重组产物稀释进含上述引物(SEQ ID NO：1和2)各0.8μM的PCR混合物中并进行由[94℃ 30秒，50℃ 30秒，72℃ 30秒]组成的循环大约15次PCR循环后，获得正确大小的单一产物(图2)。
5)克隆和分析：
用末端限制性酶BamHI和EcoO109消化上面第4步的PCR产物并按上面第2步所述进行凝胶纯化。将重组的片段连接进用BamHI和EcoO109消化的pUC18上。按厂家建议(Stratagene，San Diego CA)在标准条件下用连接混合物转化大肠杆菌并涂布到含100μg/ml氨苄青霉素，0.004％X-gal和2mM IPTG的琼脂板上。鉴别出所得的鉴定为++重组子的具有HinD III-NheI片段的菌落，因为它们是蓝色的。
本实施例说明了携带LacZα基因的1.0kb序列可消化成10-70bp的片段且这些凝胶纯化的10-70bp片段可重组进正确大小的单一产物中以便84％(N＝377)的所得的菌落是LacZ+(相对于无交换的94％；图2)。
编码来自所得LacZ-菌落的LacZ基因的DNA根据厂家说明书用测序试剂盒(美国生化公司，Cleveland OH)进行测序，发现该基因由于重组过程而具有点突变(表1)。发现11/12类型的取代且无阅读框。
                        表1
                 经诱变交换导入突变
转变          频率             易位             频率
G-A             6               A-T               1
A-G             4               A-C               2
C-T             7               C-A               1
T-C             3               C-G               0
                                G-C               3
                                G-T               2
                                T-A               1
                                T-G               2
测序了总共4,437碱基的交换LacZ DNA
从DNA测序测定10-70bp片段的DNA重组期间点诱变率为0.7％(N＝4,473)，这与PCR误差倾向相似。由于不受任何理论的限制，据信如果较大片段用于重组或如果加入防止阅读聚合酶，点诱变率可能较低。
当组合14个这些点突变的LacZ-菌落的质粒DNA并以上述方法再重组/交换时，34％(N＝291)的所得菌落是LacZ+，这些菌落推测由不同菌落的DNA重组产生。
假定诱变率为0.7％(10)，PCR误差引起的单一点突变的预期回复率可能是＜1％。
因此，可从小片段的随机混合物经过效率惊人且简单的反应重组大DNA序列。本技术的一个应用是基于同源性的相关序列的重组或交换。
实施例2.LacZ基因和完整质粒DNA杂合
1)LacZ基因的杂合
使用2个LacZ基因构建体测量以75个碱基分开两个标记之间的交换。在LacZα基因的两个分隔区插入终止编码子用作阴性标记。各标记是具有4个终止密码子的25bp非同源序列，其中两个在LacZ基因阅读框中。25bp非同源序列在图3中以大盒表示。终止密码子以盒表示或划线。含LacZα基因+-和-+型(图3)的2个1.0kb LacZ模板的1∶1的混合物用DNAseI消化并按实施例1所述纯化出100-200bp的片段。除了加入0.5μl聚合酶且总体积为100μl外在与实施例1的重组中所述相似的条件下进行交换程序。
克隆后，获得的蓝色菌落数是24％(N＝386)；它与兰色菌落的理论最大数(即，25％)相近，表明在两个标记之间完成了重组。所有10个蓝色菌落含预期的HindIII-NheI限制性片段。
2)完整质粒DNA的杂合
也试验了完整2.7kb质粒(pUC18-+和pUC18+-)。用DNA seI消化含LacZα的+-和-+型(图3)的两个2.9kb质粒的1∶1混合物并按实施例1所述纯化100-200bp的片段。除了进行[94℃ 30秒，55℃ 30秒，72℃ 30秒]的60次循环外在与上面步骤(1)重组所述相似的条件下进行交换程序。凝胶分析表明：经过交换程序后，大多数产物大于20kb。因此，完整2.7kb质粒(pUC18-+和pUC18+-)从不加引物的随机100-200bp片段中高效重组。
用在质粒上具有单一位点的限制性酶(EcoO109)消化后，大多数产物由预期大小的单一带组成。将该带凝胶纯化，再连接并将DNA用于转化大肠杆菌。转化子按实施例1所述涂布在0.004％X-gal板上。所得的质粒11％(N＝328)是兰色，因而是++重组子。
3)峰DNA杂合
混合进交换混合物中的寡核苷酸可基于该寡核苷酸侧翼序列与模板DNA的同源性掺入终产物中(图4)。上面描述的LacZ-终止密码突变子(pUC18-+)用作DNAseI消化的模板。将在两端包括与野生型LacZ基因同源的18个碱基的66聚体寡核苷酸以超出原始基因中存在的正确终止密码突变4倍的摩尔数加入到反应中。在与上面步骤2相似的条件下进行交换反应。消化所得的产物，连接后按上面所述插入大肠杆菌中。
                     表2
                                          兰菌落％
对照                                    0.0(N＞1000)
上链峰                                  8.0(N＝855)
下链峰                                  9.3(N＝620)
上、下链峰                              2.1(N＝537)
假定由于竞争性杂交，ssDNA似乎比dsDNA更有效。经过调节超出的摩尔数，退火温度，或同源性长度可大范围地改变插入程度。
实施例3.在完全缺乏引物的DNA重装配
用限制性酶EcoRI，EcoO109，XmnI和AlwNI消化质粒pUC18，产生大约370，460，770和1080bp的片段。电泳并从2％低熔点琼脂糖凝胶中分别纯化这些片段(370和460碱基的带不能分离)。产生在3个单独的试管中的大型片段，中型片段和2个小片段的混合物。
按实施例1所述用DNAseI消化各片段，对于各原始片段从2％低熔点琼脂糖凝胶中纯化50-130bp的片段。
自PCR混合物(如上面实施例1所述)中加入纯化的消化片段至片段终浓度为10ng/μl。不加入引物。分别对三个消化的DNA片段混合物进行重组反应75次循环[94℃ 30秒，60℃ 30秒]以琼脂糖电泳分析产物。
结果清楚地显示从纯化片段中有效地重新形成的1080，770和370及460bp的带，证实交换根本不需使用任何引物。
实施例4.IL-1β基因杂合
本实施例描述了可获得基于同源性不超过15个碱基的交换。作为一个例子，交换人和鼠IL-1β基因。
具有大肠杆菌密码子用法的鼠IL1-β基因(BBG49)和人IL1-β基因(BBG2；R&D Systems，Inc.Minneapolis MN)用作交换反应中的模板。人和鼠IL-1β序列的全部同源区平均仅4.1碱基长(图5，同源区用盒表示)。
各基因dsDNA PCR产物的制备，引物的清除，DNAseI消化和10-50bp片段的纯化与上面实施例1所述相似。用于PCR反应的引物序列是：5′TTAGGCACCCCAGGCTT3′(SEQ ID NO：3)和5′ATGTGCTGCAAGGCGATT3′(SEQ ID NO：4)。
用DNA聚合酶I的Klenow片段进行交换反应的前15个循环，在每次循环中加入1单位的新鲜酶。将DNA加入实施例1的PCR混合物中，该混合物没有聚合酶。操作方法是94℃ 1分钟，然后进行15次[95℃ 1分钟，干冰/乙醇上10秒(直到冷冻)，25℃保温大约20秒，加入1U的Klenow片段并在25℃保温2分钟]的循环。在每次循环中经变性步骤后，在干冰/乙醇中将试管迅速冷却并预热到退火温度。然后加入热变性聚合酶。在每次循环中都需加入该酶。使用本方法，获得了高水平的变换，其基础仅是未间断的具同源性的少数碱基(图5，交换位置以 表示)。
经过这15次循环操作后，加入Taq聚合酶，进行无引物的另外22次交换反应循环[94℃30秒，35℃30秒]。
然后将反应物稀释20倍。加入下列引物至终浓度为0.8μM：5′AACGCCGCATGCAAGCTTGGATCCTTATT3′(SEQ ID NO：5)和5′AAAGCCCTCTAGATGATTACGAATTCATAT3′(SEQ ID NO：6)，按上面实施例1所述进行PCR应。第二引物对与第一对不同，仅仅因为据信限制性位点的改变是必需的。
用XbaI和SphI消化PCR产物后，将片段连接进XbaI-SphI-消化的pUC18。用双脱氧DNA测序试剂盒(美国生化公司，Cleveland OH)根据厂家说明书测定一些菌落的插入序列。
经过对9个菌落的DNA测序发现总共17个交换。一些交换仅以未间断同源的1-2个碱基为基础。
发现为了加强以短同源性为基础的有效交换，需要非常低的有效退火温度。使用任何热稳定性聚合酶，PCR机器的冷却时间(以1-2度/秒从94℃到25℃)引起的有效退火温度比系统的退火温度更高。因此，以Taq聚合酶为基础的方法都不能产生交换，即使所使用的超出IL1-β基因的10倍。相反，热变性聚合酶，如DNA聚合酶I的Klenow片段可用于精确地获得低退火温度。
实施例5.TEM-1β内酰胺酶基因的DNA杂合
在β内酰胺酶模型系统上试验了直接分子进化的诱变DNA变换的实用性。TEM-1β内酰胺酶是一种非常有效的酶，主要以扩散限制其反应速率。本实施例确定了改变其反应特异性和获得对药品头胞噻肟的抗性是否可能，正常情况下它不水解头胞噻肟。
经过在具有不同水平的头胞噻肟(Sigma，St.Louis MO)平板上涂布10μl 10-2稀释的大肠杆菌XL1-blue细胞(Stratagene，San Diego CA)的过夜细菌培养物(大约1000cfu)确定头胞噻肟对缺乏质粒的细菌细胞的最低抑制浓度(MIC)，接着在37℃下培养24小时。
在头胞噻肟上生长对稠密的细胞具有敏感性，因此需要将相似的细胞数涂布到各板上(经过在简单LB板上涂布获得)。均匀地进行1000个细胞的涂布。
1)起始质粒构建
使用携带细菌性TEM-1β-内酰胺酶基因的pUC18衍生物(28)。TEM-1β-内酰胺酶基因赋予细菌对大约0.02μg/ml的头胞噻肟产生抗性。用下列两个引物对载体序列进行PCR：Primer A(SEQ ID NO：7)：5′TTCTATTGACGGCCTGTCAGGCCTCATATATACTTTAGATTGATTT3′andPrimer B(SEQ ID NO：8)：5′TTGACGCACTGGCCATGGTGGCCAAAAATAAACAAATAGGGGTTCCGCGCACATTT3′以及用其它如下两个引物对β内酰胺酶基因序列进行PCR：Primer C(SEQ ID NO：9)：5′AACTGACCACGGCCTGACAGGCCGGTCTGACAGTTACCAATGCTT，和Primer D(SEQ ID NO：10)：5′AACCTGTCCTGGCCACCATGGCCTAAATACATTCAAATATGTAT。
将Sfi1限制性位点加到启动子5′端和该基因3′端。
两个反应产物用SfiI消化，混合，连接并用于转化细菌。
所得的质粒是pUC182Sfi。该质粒含有携带TEM-1曲因和P-3启动子的Sfi1片段。
37℃培养24小时后头胞噻肟对携带该质粒的大肠杆菌XL1-blue(Stratagene，San Diego CA)最低抑制浓度是0.02μg/ml。
经过逐步在2倍增加的药物水平上再涂布稀释库的细胞(大约107cfu)确定无交换时β内酰胺酶基因对头胞噻肟抗性增加的能力。无交换时可获得高达1.28μG/ml的抗性。这代表抗性增加64倍。
2)DNAseI消化
第一次交换反应的底物是经过用引物C和D(两者都含SfiI位点)对pUC182Sfi进行PCR获得的0.9kb的dsDNA。
以Wizard PCR制品(Promega，Madison WI)在每次循环时去掉PCR产物的游离引物。
用在100μl 50mM Tris-HCl pH7.4，1mM MgCl2中的0.15单位的DNAseI(Sigma，St.Louis MO)在室温下消化大约5μg的DNA底物10分钟，经电泳到DE81离子交换纸(Whatman，Hillsborough OR)上从2％低熔点琼脂凝胶上纯化100-300bp的片段，以实施例1所述的方法用1M NaCl洗脱和乙醇沉淀。
3)基因杂合
将纯化片段以10-30ng/μl的浓度重悬于PCR混合物(各0.2mM的dNTP，2.2mMMgCl2，50mM KCl，10mM Tris-HCl pH9.0，0.1％Triton X-100)。此时不加引物。在MJ Research(Watertown MA)PTC-150热循环器中使用的重组方法是94℃ 60秒，然后进行40次[94℃ 30秒，50-55℃ 30秒，72℃ 30秒]的循环，再72℃ 5分钟。
4)用引物扩增重装配产物：
将重组产物稀释进含引物(C和D)各0.8μM的PCR混合物并进行20次[94℃ 30秒，50℃ 30秒，72℃ 30秒]的PCR循环后，获得900bp大小的单一产物。
5)克隆和分析：
用末端限制性酶SfiI消化900bp的产物并进行琼脂糖凝胶纯化后，用T4 DNA连接酶(BRL，Gaithersburg MD)将900bp产物连接进载体pUC182Sfi的单一SfiI位点。混合物电穿孔进大肠杆菌XL1-蓝细胞并涂布到含0.32-0.64μg/ml头胞噻肟(Sigma，St.Louis MO)的LB板上。将细胞在37℃生长24小时，从板上刮下所得的菌落作为细胞库并用作下一轮交换的PCR模板。
6)随后的重装配步骤
将3中每轮交换后获得的转化子涂布到水平增加的头胞噻肟上。合并含有最高水平头胞噻肟平板上的菌落(＞100，保持多样性)并用作下一轮PCR反应的模板。
在步骤(5)中以0.32-0.64μg/ml获得的头胞噻肟菌落的混合物用作下一轮交换的模板。10μl在LB培养基中的细胞用作重组过程的模板，重组方法是99℃ 10分钟，然后进行35次[94℃ 30秒，52℃ 30秒，72℃ 30秒]循环，再按上面所述在72℃进行5分钟。
按上面步骤(5)所述消化重组产物并连接进pUC182Sfi。混合物电穿孔进大肠杆菌XL1-blue细胞并涂布到含5-10μg/ml头胞噻肟的LB平板上。
除了细胞涂布到含80-160μg/ml头胞噻肟的LB板上外以5-10μg/ml获得的菌落用于与第一、和第二轮相似的第三轮。第三轮后，以80-160μg/ml获得菌落，在浓度增加的头胞噻肟上重新涂布后，37℃培养24小时后可获得高达320μg/ml的菌落(MIC＝320μg/ml)。
在头胞噻肟上的生长取决于细胞密度，需要所有的MICs标准化(在本例中大约1,000细胞/板)。在更高的细胞密度，获得在高达1280μg/ml时生长。在1,280μg/ml上生长的5个最大菌落涂布2次以产生单个菌落，以菌落PCR产物的限制性图谱分析Sfil插入片段。
获得了-个对头胞噻肟抗性增加16,000倍(MIC＝0.02μg/ml到MIC＝320μg/ml)的突变子。
选择后，将选定克隆的质粒转移回到野生型大肠杆菌XL1-blue细胞(Stratagene，San Diego CA)中以确保测定药物抗性的菌落不是由染色体突变引起。
三轮交换和选择产物对TEM-1β内酰胺酶扩大的广谱抗菌素头胞噻肟的最小抑制浓度增加1.6×104倍。相反，无交换的重复涂布导致抗性仅增加16倍(PCR误差或序列盒突变)。
7)序列分析
生长于1,280μg/ml的所有5个最大菌落具有与野生型TEM-1酶相同的限制性图谱。从一个这些菌落中获得的质粒的sfiI插入片段以双脱氧DNA测序(美国甘化公司，Cleveland OH)根据厂家说明书测序。所有的碱基数与修改的pBR322序列相对应(29)，氨基酸序数与ABL标准编序方法相对应(30)。以Z字母编码代表氨基酸，以一字母码表示核苷酸。术语G4205A指4205核苷酸从鸟嘌呤变成腺嘌呤。
发现9个单一碱基取代。G4205A位于β内酰胺酶P3启动子(31)-35和-10位之间。Chen和Clowes(31)获得的启动子上游突变位于这里所用的Sfil片段外，因而不能检测出来。四个突变是沉默突变(A3689G，G3713A，G3934A和T3959A)，四个导致氨基酸改变(C3448T导致Gly238Ser，A3615G产生Met182Thr，C3850T产生Glu104Lys，G4107产生Ala18Val)。
8)分子回交
然后使用与额外的野生型DNA发生的分子回交，以便去掉非义必需突变。
以上面所述的正常交换相同的方法对来自第三轮DNA交换的选定质粒进行分子回交，除了DNAseI消化和交换反应在超出40倍的野生型TEM-1基因片段存在的条件下进行外。为了使回交更有效，在交换反应中使用非常小的DNA片段(30至100bp)。回交突变子再次在含80-160μg/ml头胞噻肟(Sigma，St.Louis MO)的LB板上选择。
在超出40倍的野生型TEM-1 DNA存在的条件下用第一轮回交菌落的DNA重复该回交交换。使用小DNA片段(30-100bp)增加回交的效率。再次在含80-160μg/ml头胞噻肟的LB平板上选择第二轮回交的突变子。
将所得的转化子涂布在160μg/ml头胞噻肟上，将合并的菌落再次涂布到水平增加到1,280μg/ml的头胞噻肟上。重新涂布在1,280μg/ml获得的最大菌落以得到单一菌落。
该回交突变子抗生比野生型高32,000倍(MIC＝640μg/ml)。该突变菌株对头胞噻肟抗性比以前报道的临床或工程TEM-1来源的菌株高64倍。因此，似乎DNA交换对直接分子进化的至少一些循环而是快速且有效的工具。
使用双脱氧DNA测序试剂盒(美国生化公司，Cleveland OH)根据厂家说明书测定回交突变子SfiI插入片段的DNA序列(表3)。该突变子有9个单碱基对突变。与预期一样，所有4年以前鉴定的沉默突变丢失，回复到野生型基因的序列。在回交克隆中保留了启动子突变(G4205A)以及四个氨基酸突变中的3个(Glu104Lys，Met182Thr，和Gly238Ser)，表明它们对高水平的头胞噻肟抗性是必需的。然而，发现了2个新沉默突变(T3842C和A3767G)以及3个导致氨基酸改变的新突变(C3441T导致Arg241 His，C3886T导致Gly92Ser，G4035C导致Ala42Gly)。尽管这两个沉默突变不影响蛋白质的一级序列，但它们可能影响蛋白质的表达水平(例如以mRNA结构)和甚至可能影响蛋白质的折叠(经过改变密码子应用并从而改变中止位点，它可能涉及蛋白质的折叠)。
                      表3
               β内酰胺酶的突变
 突变类型      非回交                    回交
氨基酸改变    Ala18Lys                    -
              Glu104Lys               Glu104Lys
              Met182Thr               Met182Thr
              Gly238Ser               Gly238Ser
                 -                    Ala42Gly
                 -                    Gly92Ser
   沉默       T3959A                      -
              G3934A                      -
              G3713A                      -
              A3689G                      -
                 -           T3842C
                 -           A3767G
启动子        G4205A         G4205A
回交和非回交突变具有一个启动子突变(以其本身或组合导致表达水平增加2-3倍)以及3个普通氨基酸改变(Glu104Lys，Met182Thr和Gly238Ser)。Glu104Lys和Gly238Ser是存在于一些头胞噻肟抗性或其它TEM-1衍生物中的突变(表4)。
9)表达水平比较
经过对根据Witholt，B.(32)的方法以渗压休克制备的外周胞质提取物进行SDS-聚丙烯酰胺凝胶电泳(4-20％；Novex，San Diego CA)比较了β内酰胺酶基因在野生型质粒，非回交突变子和回交突变子中的表达水平。
纯化的TEM-1β内酰胺酶(Sigma，St.Louis MO)用作分子量标记，缺乏质粒的大肠杆菌XL1-blue细胞用作阴性对照。
突变子和回交突变子与野生型基因相比似乎产生的β内酰胺酶蛋白质率高2-3倍。启动子突变似乎导致β-内酰胺酶水平增加2-3倍。
实施例6.TEM-1β内酰胺酶基因的突变组合的构建
为了确定不同突变组合的抗生以及为了比较新突变子和公开的突变子，将一些突变子构建进相同质粒背景中。两个突变Glu104Lys和Gly238Ser称为头胞噻肟突变子。构建的所有突变子组合具有启动子突变，允许比较选定的突变子。结果如表4所示。
经PCR将特定的突变组合导入野生型pUC182Sfi中，每一突变使用2个寡核苷酸。
获得下列突变的寡核苷酸是：Ala42Gly(SEQ ID NO：11)AGTTGGGTGGACGAGTGGGTTACATCGAACT和(SEQ IDNO：12)AACCCACTCGTCCACCCAACTGATCTTCAGCAT；Gln39Lys：(SEQ ID NO：13)AGTAAAAGATGCTGAAGATAAGTTGGGTGCAC GAGTGGGTT和(SEQ ID NO：14)ACTTATCTTCAGCATCTTTTACTT；Gly92Ser：(SEQ ID NO：15)AAGAGCAACTCAGTCGCCGCATACACTATTCT和(SEQ ID NO：16)ATGGCGGCGACTGAGTTGCTCTTGCCCGGCGTCAAT；Glu104Lys：(SEQ ID NO：17)TATTCTCAGAATGACTTGGTTAAGTACTCACCAGT CACAGAA和(SEQ ID NO：18)TTAACCAAGTCATTCTGAGAAT；Met182Thr：(SEQ ID NO：19)AACGACGAGCGTGACACCACGACGCCTGTAGCAATG和(SEQ ID NO：20)TCGTGGTGTCACGCTCGTCGTT；Gly238Ser单独：(SEQ ID NO：21)TTGCTGATAAATCTGGAGCCAGTGAGCGTGGGTCTCGCGGTA和(SEQ ID NO：22)TGGCTCCAGATTTATCAGCAA；Gly238Ser and Arg241His(组合)：(SEQ ID NO：23)ATGCTCACTGGCTCCAGATTTATCAGCAAT和(SEQ ID NO：24)TCTGGAGCCAGTGAGCATGGGTCTCGCGGTATCATT；G4205A：(SEQ ID NO：25)AACCTGTCCAGGCCACCATGGCCTAAATACAATCAAATATGTATCCGCTTATGAGACAATAACCCTGATA。
从合成的寡核苷酸中凝胶纯化出这些分离的PCR片段。组合各片段10ng，重组反应在94℃进行1分钟，然后经过25次[94℃ 30秒，50℃ 30秒和72℃ 45秒]循环。在含SifI的外侧引物(来自实施例5的引物C和D)存在的条件下对重组产物进行PCR 25个循环。用SfiI消化DNA并插入野生型pUC182Sfi载体中，获得下列突变子组合(表4)。
                         表4
名称 基因型 MIC MIC来源
TEM-1 野生型 0.02
Glu104Lys 0.08 10
Gly238Ser 016 10
TEM-15 Glu104Lys/Gly238Ser* 10
TEM-3 Glu104Lys/Gly238Ser/Gln39Lys 102-32 37，15
ST-4 Glu104Lys/Gly238Ser/Met182Thr* 10
ST-1 Glu104Lys/Gly238Ser/Met182Thr/Ala18Val/T3959A/G3713A/G3934A/A3689G* 320
ST-2 Glu104Lys/Gly238Ser/Met182Thr/Ala42Gly/Gly92Ser/Arq241His/T3842C/A3767G* 640
ST-3 Glu104Lys/Gly238Ser/Met182Thr/Ala42Gly/Gly92Ser/Arq241His* 640
*所有这些突变子另外还含有G4205A启动子突变。
推断保守的突变在MIC的15个倍增体中占9个。
Glu104Lys单独显示出仅导致倍增体的MIC达0.08μg/ml，且Gly238Ser(在一些情况下具有另一个氨基酸改变)仅导致MIC为0.16μg/ml(26)。双突变子Glu104Lys/Gly238Ser具有MIC为10μg/ml。这一突变子与TE-15相对应。
这些相同的Glu104Lys和Gly238Ser突变与Glu39Lys(TEM-3)或Thr263Met(TEM-4)组合导致高水平的抗性(TEM-3为2-32μg/ml，TEM-4为8-32μg/ml)(34，35)。
含有在回交后保守的3个氨基酸改变的突变(Glu104Lys/Met182Thr/Gly238.Ser)也具有MIC为10μg/ml。这意味着各新选择的突变子加上3个已知的突变后产生的突变体导致基因对cefotaxime的抗性进一步增加32到64倍。
天然存在的，衍生自临床TEM-1的酶(TEM-1-19)各含有仅5-7个相同突变(前述)的不同组合。由于这些突变在基因中的位置完全分离，所以经过单个区域的序列盒诱变不能获得具有高cefotaxime抗性的突变子。这可以解释为什么经过标准序列盒诱变研究获得的最大MIC仅为0.64μg/ml(26)。例如，在这一研究中分别发现G104Lys和Gly238Ser突变MIC低于0.16μg/ml。使用DNA交换允许发生组合，从而发现Glu104Lys/Gly238Ser组合的MIC为10μg/ml。
本实施例的重要局限是使用单个基因作为直始点。预期如果大量相关的，天然存在的基因发生交换会发现更好的组合。存在于该混合物中的多样性比经过诱变交换产生的随机突变更有意义。例如，预期可使用来自单一物种相关基因的所有组成成份，例如预先存在的免疫系统的多样性，或从许多不同种类获得的相关基因。
实施例7.经过所有6个突变子CDRs文库的DNA杂合改进抗体A10B
A10B scFv抗体，小鼠抗-兔IgG由Pharmacia(Milwaukee WI)赠送。使用商业上可获得的Pharmacia噬菌体演示系统，它使用pCANTAB5噬菌体演示载体。
原始A10B抗体仅具有能重复的低亲合力，因为获得的克隆仅微弱地结合固相抗原(兔IgG)，(以噬菌体ELISA(Pharmacia试验试剂盒)或噬菌体滴定测量)。在竞争试验中对A10B抗体结合产生50％抑制的兔IgG浓度是13微微摩尔。观察到的低亲合力可能是由于A10B克隆的不稳定性。
根据厂家说明书测序A10B scFv DNA(美国生化公司，Cleveland OH)。根据与Kabat(33)的比较，该序列与现存抗体相似。
1)噬菌体DNA的制备：
具有A10B野生型抗体基因(10μl)的噬菌体DNA在99℃保温10分钟；然后在72℃ 2分钟。向噬菌体DNA中加入PCR混合物(50mM KCl，10mM Tris-HClpH9.0，0.1％Triton X-100，各200μm的dNTP，1.9mM MgCl2)，各0.6μm的引物和0.5μm Taq DNA聚合酶(Promega，Madison WI)。PCR程序进行了35个[94℃30秒，45℃ 30秒，72℃ 45秒]循环。使用的引物是：5′ATGATTACGCCAAGCTTT3′(SEQ ID NO：26)和5′TTGTCGTCTTTCCAGACGTT3′(SEQ ID NO：27)。
然后电泳850bp PCR产物并从2％低熔点琼脂糖凝胶中纯化。
2)片段化
将300ng凝胶纯化的850bp带用存在于50mM Tris-HCl pH7.5，10mM MgCl2中的0.18单位的DNAseI(Sigma，St.Louis MO)室温消化20分钟。消化的DNA在2％低熔点琼脂凝胶上分离并从凝胶中纯化出50至200bp之间的带。
3)试验文库的构建
本实验的目的是试验CDRs的插入是否有效。
合成下列具有内部限制性酶位点的CDR序列。“CDR H”指重链中的CDR，“CDR L”指抗体中轻链上的CDR。
具有限制性位点的CDR寡聚体：CDR H1(SEQ ID NO：34)5′TTCTGGCTACATCTTCACAGAATTCATCTAGATTTGGGTGAGGCAGACGCCTGAA3′CDR H2(SEQ ID NO：35)5′ACAGGGACTTGAGTGGATTGGAATCACAGTCAAGCTTATCCTTTATCTCAGGTCTCGAGTTCCAAGTACTTAAAGGGCCACACTGAGTGTA3′CDR H3(SEQ ID NO：36)5′TGTCTATTTCTGTGCTAGATCTTGACTGCAGTCTTATACGAGGATCCATTGGGGCCAAGGGACCAGGTCA3′CDR L1(SEQ ID NO：37)5′AGAGGGTCACCATGACCTGCGGACGTCTTTAAGCGATCGGGCTGATGGCCTGGTACCAACAGAAGCCTGGAT3′CDR L2(SEQ ID NO：38)5′TCCCCCAGACTCCTGATTTATTAAGGGAGATCTAAACAGCTGTTGGTCCCTTTTCGCTTCAGT3′CDR L3(SEQ ID NO：39)5′ATGCTGCCACTTATTACTGCTTCTGCGCGCTTAAAGGATATCTTCATTTCGGAGGGGGGACCAAGCT3′
将CDR寡聚体以超出10倍的摩尔数加入上面步骤(2)50至200bp之间的纯化A10B抗体DNA片段中。加入PCR混合物(50mM KCl，10mM Tris-HCl pH9.0，0.1％Triton X-100，1.0mM MgCl2各200μm的dNTP，0.3μl Taq DNA聚合酶(Promega，Madison WI)，总体积为50μl)，交换程序是在94℃进行1分钟，在72℃进行1分钟，然后进行35次循环：94℃ 30秒，55℃ 30秒，72℃ 30秒。
将1μl交换混合物加入100μl PCR混合物(50mM KCl，10mM Tris-HCl pH9.0，0.1％Triton X-100，各200μm的dNTP，1.9mM MgCl2，各0.6μm的两种外侧引物(SEQ ID NO：26和27，见下面)，0.5μl Taq DNA聚合酶)中，PCR程序进行30个[94℃ 30秒，45℃ 30秒，72℃ 45秒]循环。所得的850碱基对大小的DNA片段混合物用酚/氯仿提取并用乙醇沉淀。
外侧引物是：
外侧引物1：SEQ ID NO：27
5′TTGTCGTCTTTCCAGACGTT3′
外侧引物2：SEQ ID NO：26
5′ATGATTACGCCAAGCTTT3′
将850bp PCR产物用限制性酶SfiI和NotI消化，从低熔点琼脂糖凝胶中纯化并连接进从Pharmacia，Milwankee WI获得的pCANTAB5表达载体中。根据Invitrogen(San Diego CA)阐述的方法将连接的载体电穿孔进TG1细胞(Pharmacia，MilwallKee WI)并涂布成单个菌落。
将所得菌落的DNA加入100μl PCR混合物(50mM KCl，10mM Tris-HCl，pH9.0，0.1％Triton X-100，各200μm的dNTP，1.9mM MgCl2，0.6μm的外侧引物1(SEQ ID NO：27；见下面)，6个内部引物(SEQ ID NO：40-45；见下面)，和0.5μl Taq DNA聚合酶)中，PCR过程进行35次[94℃ 30秒，45℃ 30秒，72℃45秒]循环。以琼脂糖凝胶电泳测定PCR产物的大小并用于确定哪一个具有限制性位点的CDRs被插入。CDR内部引物：H1(SEQ ID NO：40)5′AGAATTCATCTAGATTTG3′H2(SEQ ID NO：41)5′GCTTATCCTTTATCTCAGGTC3′H3(SEQ ID NO：42)5′ACTGCAGTCTTATACGAGGAT3′L1(SEQ ID NO：43)5′GACGTCTTTAAGCGATCG3′，L2(SEQ ID NO：44)5′TAAGGGAGATCTAAACAG3′，L3(SEQ ID NO：45)5′TCTGCGCGCTTAAAGGAT3′
在野生型A10B抗体DNA的预期位置插入6个合成的CDRs(图7)。这些研究表明：尽管特定克隆的6个CDRs中的每一个成为具有限制性位点的CDR的机会较小，但大多数克隆携带至少一个具有限制性位点的CDR，并产生了具有限制性位点的CDR的任意可能的组合。
4)突变的互补决定区(“CDRs”)的构建：
根据我们的序列资料，制备了与6个CDRs相应的6个寡核苷酸。CDR(Kabat定义)在合成时突变的比率为70(现存碱基)：10∶10∶10，以大约20碱基的侧翼序列在5′和3′两侧，当以摩尔数超出的量混合进未诱变的抗体基因片段混合物中时它为CDR的插入提供了同源性。所得的突变序列在下面给出：
CDR文库的寡核苷酸：CDR H1(SEQ ID NO：28)5′TTCTGGCTACATCTTCACAACTTATGATATAGACTGGGTGAGGCAGACGCCTGAA3′CDR H2(SEQ ID NO：29)5′ACAGGGACTTGAGTGGATTGGATGGATTTTTCCTGGAGAGGGTGGTACTGAATACAATGAGAAGTTCAAGGGCAGGGCCACACTGAGTGTA3′CDR H3(SEQ ID NO：30)5′TGTCTATTTCTGTGCTAGAGGGGACTACTATAGGCGCTACTTTGACTTGTGGGGCCAAGGGACCACGGTCA3′CDR L1(SEQ ID NO：31)5′AGAGGGTCACCATGACCTGCAGTGCCAGCTCAGGTATACGTTACATATATTGGTACCAACAGAAGCCTGGAT3′CDR L2(SEQ ID NO：32)5′TCCCCCAGACTCCTGATTTATGACACATCCAACGTGGCTCCTGGAGTCCCTTTTCGCTTCAGT3′CDR L3(SEQ ID NO：33)5′ATGCTGCCACTTATTACTTGCC AGGAGTGGAGTGGTTATCCGTACACGTTCGGAGGGGGGACCAAGC T3′
黑体字及划线序列是使用核苷酸70∶10∶10∶10的混合物合成的突变序列，其中70％是野生型核苷。
将摩尔数超出10倍的CDR突变寡核苷酸加入以上面步骤(2)得到的长度在50至200bp之间的纯化A10B抗体DNA片段中。加入PCR混合物(50mM KCl，10mMTris-HCl pH9.0，0.1％Triton X-100，1.9mM MgCl2，各200μm dNTP，0.3μl TaqDNA聚合酶(Promega，Madison WI)，总体积为50μl)，交换程序在94℃进行1分钟，72℃ 1分钟，然后进行35个[94℃ 30秒，55℃ 30秒，72℃ 30秒]循环。
将1μl交换混合物加入100μl PCR混合物(50mM KCl，10mM Tris-HClpH9.0，0.1％Triton X-100，各200μm的dNTP，1.9mM MgCl2，各0.6μm的两个外侧引物(SEQ ID NO：26和27，见下面)，0.5μl Taq DNA聚合酶)中，PCR程序进行30个[94℃ 30秒，45℃ 30秒，72℃ 45秒]循环，所得的850碱基对大小的DNA片段混合物用酚/氯仿提取并用乙醇沉淀。
外侧引物是：外侧引物1：SEQ ID NO：27 5′TTGTCGTCTTTCCAGACGTT3′外侧引物2：SEQ DI NO：26 5′ATGATTACGCCAAGCTTT3′
5)将scFv抗体DNA克隆进pCANTAB5
用限制性酶SfiI和NotI消化850bp PCR产物。从低熔点琼脂糖凝胶中纯化并连接进从Pharmacia，Milwaukee WI获得的pCANTAB5表达载体中。根据Invitrogen(San Diego CA)阐述的方法将连接的载体电穿孔进TG1细胞(Pharmacia，Milwaukee WI)，按照厂家建议的说明书使用辅助噬菌体生长噬菌体文库。
使用6轮选择筛选在该方法产生的文库中改进抗体的存在。
6)高亲合力克隆的选择
用兔IgG(Jackson Immunoresearch，Bar Harbor ME)以10μg/孔在37℃覆盖96孔微8滴板的15个孔1小时，然后用2％的溶于PBS的脱脂半乳在37℃封闭1小时。
用100μl 2％乳液在室温下封闭100μl的噬菌体文库(1×1010cfu)30分钟，然后加到15个孔的各孔中并在37℃保温1小时。
然后用含0.5％Tween-20的PBS洗涤各孔3次，每次10分钟。结合的噬菌体用100μl洗脱缓冲液(甘氨酸-HCl，pH2.2)洗脱，接着用2M Tris pH7.4立即中和并转染用于噬菌体生产。重复该选择循环6次。
第6个循环后，挑出单个噬菌体克隆并以噬菌体ELISA比较相对亲合力，用Pharmacia(Milwaukee WI)的试剂盒根据厂家建议的方法试验兔IgG的特异性。
当以Western分析试验时，与野生型A10B比较，最好的克隆表达水平提高大约100倍。与最佳克隆在竞争试验中产生50％抑制的兔IgG浓度是1微微摩尔。最佳克隆对兔抗原具有可重复的特异性。以噬菌体显示的抗体拷贝数似乎增加。
实施例8.经部分基因的直接重复进行体内重组
以两部分灭活的相同基因(β-内酰胺酶)拷贝构建质粒以证实这两个直接重复的共同区之间的重组产生全长的，有活性的重组基因。
使用携带细菌TEM-1β内酰胺酶基因的pUC18衍生物(Yanish-Perron et al.，1985，Gene 33：103-119)。TEM-1β内酰胺酶基因(“Bla”)赋予细菌对大约0.02μg/ml的cefotaxime产生抗性。用2个引物对载体序列进行PCR：Primer A(SEQ ID NO：46)5′TTCTATTGACGGCCTGTCAGGCCTCATATATACTTTAGATTGTTTT3′Primer B (SEQ ID NO：47)5′TTGACGCACTGGCCATGGTGGCCAAAAATAAACAAATAGGGGTTCCGCGCACATTT3′
及用另外两个引物对β-内酰胺酶基因序列进行PCR：Primer C(SEQ ID NO：48)5′AACTGACCACGGCCTGACAGGCCGGTCTGACAGTTACCAATGCTT3′Primer D(SEQ ID NO：49)5′AACCTGTCCTGGCCACCATGGCCTAAATACATTCAAATATGTAT3′
将Sfil限制性位点加到启动子5′端和β内酰胺酶基因3′端。
用Sfil消化两个反应产物，混合，连接并用于以下述程序转化受体大肠杆菌。所得的质粒是pUC182Sfi-Bla-Sfi。该质粒含有携带Bla基因和P-3启动子的Sfil片段。
携带pUC182Sfi-Bla-Sfi的大肠杆菌XL1-blue(Stratagene，San Diego CA)在37℃培养24小时后的cefotaxime最低抑制浓度为0.02μg/ml。
使用同源区域将pBR322的四环素基因克隆区pUC18Sif-Bla-Sfi，产生pBR322TetSfi-Bla-Sfi。然后用SspI和FspI限制性消化pBR322 TetSfi-Bla-Sfi去掉TEM-1基因并连接末端，产生pUC322 TetSfi-Sfi。
使用标准PCR技术和下列引物扩增TEM-1基因的重叠区：Primer 2650(SEQ ID NO：50)5′TTCTTAGACGTCAGGTGGCACTT3′Primer 2493(SEQ ID NO：51)5′TTT TAA ATC AAT CTA AAG TAT3′Primer 2651(SEQ ID NO：52)5′TGCTCATCCACGAGTGTGGAGAAGTGGTCCTGCAACTTTAT3′，以及rimer 2652(SEQ ID NO：53)ACCACTTCTCCACACTCGTGGATGAGCACTTTTAAAGTT
用Sfil和BstX1消化2个所得的DNA片段并连接进pBR322 Tet Sfi-Sfi的Sfi位点。所得的质粒称为pBR322Sfi-BL-LA-Sfi。质粒的图谱以及功能性β-内酰胺酶的质粒内重组和再构建示意图在图9中表示。
将质粒电穿孔进TG-1或JC8679大肠杆菌细胞。大肠杆菌JC8679是RecBC sbcA(Dliner et al.，1993，NAR 21：5192)。将细胞涂布到含四环素的琼脂平板上。然后将生长的那些克隆涂布到含100μg/ml氨苄青霉素的固体琼脂板上并计数活菌落数。将表现出氨苄青霉素抗性的那些转化子中的β内酰胺酶基因插入片段以标准PCR技术扩增，使用的引物是：Primer 2650(SEQ ID NO：50)5′TTCTTAGACGTCAGGTGGCACTT3′和Primer 2493(SEQ ID NO：51)5′TTTTAAATCAATCTAAAGTAT3′
测量插入片段的长度。1kb插入片段的存在表明该基因已成功地重组，如图9和表5所示。
                      表5
    细胞     菌落    菌落        菌落
    TG-1     131     21     3/3 at 1 kb
    JC8679     123     31     4/4 at 1 kb
    载体对照     51     0
大约17-25％的四环素抗性菌落也是氨苄青霉素抗性，经菌落PCR测定全部氨苄青霉素抗性菌落已正确重组。因此，位于相同质粒的部分基因将成功地重组以产生功能基因。
实施例9.经全长基因的直接重复进行体内重组
构建具有β内酰胺酶基因不同等位基因的两个全长拷贝的质粒。两个基因的同源重组产生该基因的单个重组全长拷贝。
pBR322Tet Sfi-Sfi和pBR322 Tet Sfi-Bla-Sfi的构建描述如上。
按下面构建β内酰胺酶基因的两上等位基因。用pUC18Sfi-Bla-Sfi作模板进行两个PCR反应。一个反应用下列引物进行：Primer 2650(SEQ ID NO：50)5′TTCTTAGACGTCAGGTGGCACTT3′Primer 2649(SEQ ID NO：51)5′ATGGTAGTCCACGAGTGTGGTAGTGACAGGCCGGTCTGACAGTTACCAATGCTT3′
用下列引物进行第二个PCR反应：Primer 2648(SEQ ID NO：54)5′TGTCACTACCACACTCGTGGACTACCATGGCCTAAATACATTCAAATATGTAT3′Primer 2493(SEQ ID NO：51)5′TTT TAA ATC AAT CTA AAG TAT 3′
这样产生两个Bla基因，一个具有5′Sfi1位点和3′BstX1位点，另一个具有5′Bst X1位点和3′Sfi1位点。
用BstX1和Sfi1消化这两个基因后，连接进Sfi1消化的质粒pBR322Tet Sfi-Sfi中，获得具有Bla基因串联重复的质粒(pBR322-Sfi-2BLA-Sfi)，(见图10)。将该质粒电穿孔进大肠杆菌细胞中。将细胞涂布到含15μg/ml四环素的固态琼脂板上。然后将那些生长的菌落涂布到含100μg/ml氨苄青霉素的固态琼脂板上并计数活菌落数。将表现出氨苄青霉素抗性的转化子中的Bla插入片段以标准PCR技术使用实施例8描述的方法和引物扩增。1kb插入片段的存在表明：复制基因已重组如表6所示。
                      表6
    细胞     菌落    菌落       菌落
    TG-1      28     54     7/7 at 1kb
    JC8679     149    117     3/3 at 1kb
    载体对照      51      0
菌落PCR证实串联重复有效重组形成了单个重组基因。
实施例10.多轮直接重复重组-质粒内
为了确定多轮重组是否可用于更快地生产抗性细胞，进行实施例9所述方法的多个循环。
负重组控制由单拷贝的β内酰胺酶基因组成，而正重组实验由插入的两个拷贝的β内酰胺酶直接重复组成。四环素标记用于平衡在每轮中选择cefotaxime抗性的菌落数以补偿连接效率。
在第一轮中，用EcrI消化pBR322TetSfi-Bla-Sfi并用正常和Cadwell PCR混合物(Cadwell and Joyce(1992)PCR Methods and Applications 2：28-33)的1∶1混合物(1ml)进行PCR以获得发生误差的PCR。PCR程序是开始在70℃ 2分钟。然后进行30次循环，每循环在94℃ 30秒；52℃ 30秒和72℃ 3分钟6秒，接着在72℃ 10分钟。
在PCR反应中用于产生一个Bla基因控制质粒的引物是引物2650(SEQ IDNO：5))和引物2719(SEQ ID NO：55)5′TTAAGGGATTTTGGTCATGAGATT3′。这产生扩增DNA片段的混合群体，集中命名为片段#59。这些片段有不同突变数目。
用于两个不同PCR反应以产生两个Bla基因质粒的引物对于第一基因是引物2650(SEQ ID NO：50)和引物2649(SEQ ID NO：51)，对于第二基因是引物2719(SEQID NO：55)。这产生两个扩增DNA片段中每一个的混合群体：片段#89(用引物2648和2719扩增)和片段#90(用引物2650和2649扩增)。在每种情况下将不同突变数导入各片段的混合群体中。
经过有误差的PCR后，用Sfil消化扩增DNA片段#59的群体，然后克隆进pBR322TetSfi-Sfi以产生质粒pBR322Sfi-Bla-Sfi1的混合群体。
经过有误差的PCR后，用SfiI和BstXI在50℃消化扩增DNA片段#90和#89的群体并连接进pBR322TetSfi-Sfi以产生质粒pBR322TetSfi-2Bla-Sfi1的混合群体(图10)。
将质粒pBR322Sfi-Bla-Sfi1和pBR322Sfi-2Bla-Sfi1电穿孔进大肠杆菌JC8679并涂布在具有不同cefotaxime浓度的琼脂板上以选择抗性菌株并涂布在四环素板上滴定。
挑出等数目的菌落(根据在四环素上生长的菌落数)，在LB-tet上生长并从菌落中提取DNA。这是一轮重组。用EcrI消化该DNA并按上面所述用于第二轮有误差的PCR。
经过5轮后，一个片段质粒对cefotaxime的MIC(最低抑制浓度)是0.32，而两个片段质粒的MIC是1.28。结果表明，经过5次循环后，重组获得的抗性比体内重组存在的抗性高4倍。
实施例11.经片段的电穿孔进行体内重组
按所述制备含pUC18Sfi-Bla-Sfi的感受态大肠杆菌。如上所述质粒pUC18Sfi-Bla-Sfi含标准TEM-1β-内酰胺酶基因。
获得来自pUC18Sfi-cef-Sfi(克隆ST2)(Stemmer WPC(1994))Nature370：389-91，本文引用作为参考)的TE-1衍生的cefotaxime抗性基因，它赋予携带该质粒的大肠杆菌对cefotaxime的MIC为640μg/ml。在一个实验中，将完整的质粒pUC18Sfi-cef-Sfi DNA电穿孔进具有质粒pUC18Sfi-Bla-Sfi的大肠杆菌细胞中。
在另一实验中，将含来自pUC18Sfi-cef-Sfi的cefotaxime基因的DNA片段使用引物2650(SEQ ID NO：50)和2719(SEQ ID NO：55)经PCR扩增。所得的1kb PCR产物以DNase消化成＜100bp的DNA片段，将这些片段电穿孔进已含有pUC18Sfi-Bla-Sfi的感受态大肠杆菌细胞中。
然后经过将转化细胞涂布到具有不同cefotaxime浓度的琼脂板上试验来自两个实验中的转化细胞对cefotaxime的抗性。结果在表7中表示。
                      表7  菌落/cefotaxime浓度
无DNA对照     0.16    0.32    1.28     5.0     10.0
ST-2突变子，整体      14
ST-2突变子，片段    4000    2000     800     400
野生型，整体    1000     120     22     7
野生型，片段      27
     18
该结果表明电穿孔后完整ST-2 Cef基因插入了细菌基因组成质粒中。由于大多数插入片段是同源的，预期该基因插入质粒中，取代野生型基因。St-2的Cef基因片段也有效地插入质粒的野生型基因中。对导入野生型基因(整体或片段)和无DNA观察不到cefotaxime抗性的急剧增加。因此，St-2片段显示出比野生型片段产生更大的cefotaxime抗性。期望从增加抗性基因库中制备的重复的插入片段会导致抗性增加。
因此，分离具有St-2基因片段，产生增加的cefotaxime抗性的那些菌落并提取质粒DNA。使用上面描述的PCR方法扩增该DNA。用DNase将扩增DNA消化成片段(＜100bp)，将2-4μg该片段电穿孔进已含有pUC322Sfi-Bla-Sfi的上面所述的感受态大肠杆菌细胞中。将转化的细胞涂布在含有不同cefotaxime浓度的琼脂上。
作为对照，也使用具有质粒pUC18Sfi-Kan-Sfi的感受态大肠杆菌。将pUC18Sfi-cef-Sfi PCR产物消化的DNA片段电穿孔进这些细胞中。在卡那霉素基因和β内酰胺酶基因间无同源性，因此不会发生重组。
本实验重复2轮，结果在表8中显示。
                     表8
次数 浓度 对照 抗性菌落
1重复涂布 0.16-0.640.32 菌落10小 菌落1000
2重复涂布 10 10100sm@2.5 40050@10
3 401280 100sm 100sm
实施例12.重组形式的确定
设计本实验以确定哪一种重组形式在每个循环中产生最多重组子。
在第一种方法中，用PCR引物扩增载体pUC18Sfi-Bla-Sfi以产生大型和小型片段。大片段具有质粒和部分Bla基因的末端，小片段编码中央Bla基因。使用实施例8的PCR方法产生具有完整Bla基因的第三个片段。以上面描述的方法将大质粒片段和含完整Bla基因的片段同时电穿孔进大肠杆菌JC8679细胞中，将转化子涂布在不同浓度的cefotaxime上。
在第二种方法中，扩增载体pUC18Sfi-Bla-Sfi以产生如上面第一种方法中分离的大质粒片段。经PCR获得各包含部分完整Bla基因的2个片段，使两个片段一起覆盖完整的Bla基因。将大质粒片段和2个Bla基因片段都电穿孔进感受态的大肠杆菌JC8679细胞中，将转化子涂布在不同浓度的cefotaxime上。
在第三种方法中，将载体和质粒都电穿孔进大肠杆菌JC8679细胞中并将转化子涂布在不同浓度的cefotaxime上。
在第四种方法中，将完整Bla基因电穿孔进已含有载体pUCSfi-Sfi的大肠杆菌细胞保并将转化子涂布在不同浓度的cefotaxime上。作为对照用完整Bla基因或仅用载体电穿孔大肠杆菌JC8671细胞。
结果在图11中表示，与使用具有完整Bla基因的一个片段相比，两个片段插入载体的效率低100倍。第3步研究表明：插入效率取决于自由DNA末端的存在，因为在这一步研究中设有获得重组子。然而，方法3中的结果也是由于载体电穿孔的效率低，当表达载体已存在于感受态细胞中时，载体电穿孔的效率不再是因素，甚至用未切割的载体可获得有效的同源重组。
实施例13.优化载体效果的序列盒杂合试剂盒
为了提供能赋予优化表型的载体(例如，载体编码序列的最大表达，如克隆的基因)，提供了一套试剂盒，包含各种能被交换的序列盒，并选择优化的交换子。图12是一个实施方案的图示，每个基因座具有许多序列盒。例如，在细菌表达系统中，图13显示了在各自基因座使用的序列盒例子。给字基因座的各序列盒(例如，本实施例中的所有启动子)两侧是能重叠邻近基因座序列盒侧翼序列且优选也能参与同源重组或非同源重组(例如，lox/cre或flp/frt系统)的基本相同的序列，以便实现在一个基因座内而基本上不是基因座之间的序列盒的交换。
在试剂盒中以PCR片段的形式提供序列盒，其中各序列盒类型或单个序列盒种类包装在分离的试管中，经过合并试管的内容物以装配完整质粒或其主要部分构建载体文库，其中经过使每个基因座序列盒重叠的侧翼序列与邻近基因座的序列盒杂交来实现。装配的载体连接到预定的目的基因上以形成载体文库，其中各文库成员包含预定的目的基因和序列盒的组合，这经过序列盒的联接来确定。将载体转移进合适的宿主细胞中，在适合于表达的条件下培养细胞并选择所需的表型。
尽管使用认为是优选的实施例对本发明进行了描述，但应清楚本发明不受所公开实施例的限制。相反，本发明试图覆盖包括在所附权利要求的实质和范围内的各种修改和等价方法。
                      参考文献
本申请在说明书的相关部分引用了下列参考文献：
1.J·H·霍兰德，《美国科学》，1992，7，66-72。
2.J·H·霍兰德，《天然和人工系统的适应》，1992，第二版，MIT出版，剑桥。
3.G·F·乔易斯，《科学美国人》，1992，12，90-97。
4.S·A·考夫曼，《秩序起源》，1993，牛津大学出版，纽约。
5.G·D·斯多莫，《酶学方法》，1991，208：458-468。
6.T·D·施奈德等，《分子生物学杂志》，1988，188：415-431。
7.J·F·瑞德哈尔奥森和R.T.绍尔，《科学》，1988，241：53-57。
8.W·P·C·斯提莫等，《生物技术》，1992，14：256-265。
9.H·P·约克等，《理论生物学杂志》，1977，67：345-376。
10.H·P·约克等，《理论生物学杂志》，1974，46：369-380。
11.D·W·利昂等，《技术》，1989，1：11-15。
12.R·C·卡尔德威尔和G·F乔易斯，《PCR方法和应用》，1992，2：28-33。
13.D·P·巴特尔和J·W·佐斯泰克，《科学》，1993，261：1411-1418。
14.L·C·博克等，《自然》，1992，355：564-566。
15.J·K·斯各特和G.P.史密斯，《科学》，1990，249：386-390。
16.S·E·克威尔拉等，《美国自然科学院进展》，1990，87：6378-6382。
17.S·E·麦克卡佛蒂等，《自然》，1990，348：552-554。
18.M·G·卡尔等，《美国自然科学院进展》，1992，89：1865～1869。
19.H·格拉姆等，《美国自然科学院进展》，1992，89：3576-3580。
20.A·阿金和D·C·尤文，《美国自然科学院进展》，1992，89：7811-7815。
21.A·R·奥利芬特等，《基因》，1986，44：177-183。
22.J·D·赫米斯等，《美国自然科学院进展》，1990，87：696-700。
23.A·迈尔汉斯等，《核酸研究》，1990，18：1687-1691。
24.J·J·奥斯特霍特等，《美国化学社会杂志》，1992，114：331-337。
25.R·J·卡农等，《自然》，1993，363：536-538。
26.帕尔兹基尔和博特斯蒂思，《细菌学杂志》，1992，174：5237-5243。
27.马腾等，《核酸研究》，19：2423。
28.杨尼斯-佩龙等，《基因》，1985，33：103-119。
29.瓦特森，《基因》，1988，70：399-403。
30.安布勒等，《生物化学杂志》，1991，276：269-272。
31.切恩和克洛斯，《核酸研究》，1984，12：3219-3234。
32.B·威特赫特，《分析生物化学》，1987，164(2)：320-330。
33.卡巴特等，“免疫学蛋白质的序列”美国健康与人类服务部，NIH发表，19-3242。
34.菲利蓬等，《抗微生物试剂化学》，1989，33：1131-1136。
35.雅各比和梅代罗斯，《抗微生物试剂化学》，1991，35：167～1704。
36.科埃略萨姆陪奥，《生物化学》，1993，32：10929-10935。
37.C·图尔克等，《美国自然科学院进展》，1992，89：6988-6992。
38.第4,683,195号美国专利。
39.第4,683,202号美国专利。
40.德拉哥瑞伍等，《蛋白质工程》，1993，6：327-331。
41.德拉哥瑞伍等，《生物/技术》，1993，11：1548-1552。
42.ER.戈德曼和DC.优文，《生物/技术》，1992，10：1557-1561。
43.尼斯姆等，《EMBO杂志》，1994，13：692-698。
44.温特等，《免疫学年志》，1994，12：433-55。
45.凯伦等，《生物/技术》，1994，12：517-520。
46.卡洛盖洛等，《FEMS微生物学通信》，1992，97：41-44。
47.盖利兹等，WO91/01087。
48.海雅西等，《生物技术》，1994，17：310-315。
49.拉达姆等，WO90/07576。(1. Holland，J.H.(1992)Sci.Am.July，66-72.2. Holland，J.H.(1992)″Adaptation in natural and artificial
systems″Second edition，MIT Press，Cambridge.3. Joyce，G.F.(1992)Scientific American，December，90-97.4. Kauffman，S.A.(1993)″The origins of order″.Oxford
University Press，New York.5. Stormo，G.D.(1991)Methods Enzymol.208：458-468.6. Schneider，T.D.et al.，(1986)J.Mol.Biol.188：415-431.7. Reidhaar-Olson，J.F and Sauer，R.T.(1988)Science 241：53-
57.8. Stemmer，W.P.C.et al.，(1992)Biotechniques 14：256-265.9. Yockey，H.P.(1977)J.Theor.Biol.67：345-376.10.Yockey，H.P.(1974)J.Theor.Biol.46：369-380.11.Leung，D.W.et al.，(1989)Tachnique 1：11-15.12.Caldwell，R.C.and Joyce，G.F.(1992)PCR Methods and
Applications 2：28-33.13.Bartel，D.P.，and Szostak，J.W.(1993)Science 261：1411-1418.14.Bock，L.C.et al.，(1992)Nature 355：564-566.15.Scott，J.K.and Smith，G.P.(1990) Science 249：386-390.16.Cwirla，S.E.et al.(1990)Proc.Natl.Acad.Sci.USA87：6378-6382.17.McCafferty，J.et al.(1990)Nature 348：552-554.18.Cull，M.G.et al.，(1992)Proc.Natl.Acad.Sci.USA89：1865-1869.19.Gramm，H.et al.，(1992)Proc. Natl.Acad.Sci.USA 89：3576-3580.20.Arkin，A.and Youvan，D.C.(1992)Proc.Natl.Acad.Sci.USA 89：7811-7815.21.Oliphant，A.R.et al.，(1986)Gene 44：177-183.22.Hermes，J.D.et al.，(1990)Proc.Natl.Acad.Sci.USA87：696-700.23.Meyerhans，A.et al.，(1990)Nucleic Acids Res.18：1687-1691.24.Osterhout，J.J.et al.，(1992)J.Am.Chem.Soc.114：331-337.25.Cano，R.J.et al.，(1993)Nature 363：536-538.26.Palzkill and Botstein，(1992)J.Bacteriol.174：5237-5243.27.Marton et al.，Nucleic Acids Res.19：2423.28.Yanish-Perron et al.，[1985] Gene 33：103-119.29.Watson(1988)Gene 70：399-403.30.Ambler et al.(1991)Biochem J.276：269-272.31.Chen and Clowes，(1984)Nucleic Acid Res.12：3219-3234.32.Witholt，B.([1987] Anal.Biochem.164(2)：320-33033.Kabat et al.，(1991)″Sequences of Proteins of ImmunologicalInterest″U.S.Department of Health and Human Services，NIHPublication 91-3242.34.Philippon et al.，(1989)Antimicrob Agents Chemother33：1131-1136.35.Jacoby and Medeiros(1991)Antimicrob.Agents Chemother.35：167-1704.36.Coelhosampaio(1993)Biochem.32：10929-1093537.Tuerk，C.et al.，(1992)Proc.Natl.Acad.Sci.USA 89：6988-6992.38.United States Patent No.4,683,19539.United States Patent No.4,683,20240.Delagrave et al.(1993)Protein Engineering 6：327-33141.Delgrave et al.(1993)Bio/Technology 11：1548-155242.Goldman，ER and Youvan DC(1992)Bio/Technology 10：1557-156143.Nissim et al.(1994)EMBO J.13：692-69844.Winter et al.(1994)Ann.Rev.Immunol.12：433-5545.Caren et al.(1994)Bio/Technology 12：517-52046.Calogero et al.(1992)FEMS Microbiology Lett.97：41-4447.Galizzi et al. WO91/0108748.Hayashi et al.(1994)Biotechniques 17：310-31549.Radman et al.WO90/07576)
此处引用的所有文献，专利和专利申请以其整体所为参考，其程度与以其全文作为参考引用时具体和单独说明的各文献、专利或专利申请一样。

Claims (32)

1.用于将一个或多个突变导入模板双链多核苷酸的方法，其中模板双链多核苷酸已裂解成所需大小的双链随机片段，该方法包括：
a)将一种或多种单链或双链寡核苷酸加入所得的双链片段群体中，其中所说的寡核苷酸包含与模板多核苷酸相同的区域和异源区域；
b)将所得的双链随机片段与寡核苷酸的混合物变性成单链片段；
c)在导致所说单链片段在单链片段之间相同区域上退火的条件下将所得的单链片段群体与聚合酶保温并形成诱变的双链多核苷酸；以及
d)重复步骤(b)和(c)。
2.权利要求1的方法，其中在双链片段混合物中特定双链片段的浓度不超过总DNA重量的1％。
3.权利要求1的方法，其中不同特定双链片段的数目包括至少大约100个。
4.权利要求1的方法，其中双链片段的大小从56bp到5kb。
5.权利要求1的方法，其中诱变的双链多核苷酸的大小包含50bp到100kb。
6.生产具有生物活性重组蛋白质的方法，包括：
a)在提供将所说模板多核苷酸裂解成具有所需大小的随机双链片段的条件下处理包含编码野生型蛋白质的双链模板多核苷酸的样品；
b)向所得的随机片段的群体中加入一种或多种单链或双链寡核苷酸，其中所说的寡核苷酸包含模板多核苷酸的相同区和异源区；
c)将所得的双链随机片段和寡核苷酸的混合物变性成单链片段；
d)在导致所说的单链片段在相同区域退火的条件下将所得的单链片段群体与聚合酶保温并形成诱变的双链多核苷酸；
e)重复步骤(c)和(d)；
f)从诱变的双链多核苷酸表达重组蛋白质。
7.权利要求6的方法，其中在步骤(a)中双链片段混合物中的特定双链片段的浓度不到总DNA重量的1％。
8.权利要求6的方法，其中步骤(a)中不同的特定双链片段的数目包含至少大约100个。
9.权利要求6的方法，其中双链片段的大小是从大约5bp到5kb。
10.权利要求6的方法，其中诱变的双链多核苷酸大小包括从50bp到100kb。
11.权利要求6的方法，进一步包含从重组蛋白质群体中选择所需的重组蛋白质。
12.获得嵌合多核苷酸的方法，包含：
a)在提供将所说的模板多核苷酸裂解成所需大小的随机双链片段的条件下处理包含不同双链模板多核苷酸的样品，其中所说的不同模板多核苷酸含有相同区和异源区；
b)将包含在步骤(a)产生的处理样品中的所得随机双链模板片段变性成单链片段；
c)在提供靶单链片段在相同区域退火的条件下将所得的单链片段与聚合酶保温并形成包含模板多核苷酸序列的嵌合双链多核苷酸序列；
d)按需要重复步骤(b)和(c)。
13.权利要求12的方法，其中在步骤(a)中双链片段混合物中的特定双链片段浓度不超过总DNA重量的1％。
14.权利要求12的方法，其中步骤(a)不同的特定双链片段的数目包含至少大约100个。
15.权利要求12的方法，其中双链片段的大小是从大约5bp到5kb。
16.权利要求12的方法，其中诱变的双链多核苷酸的大小包含从50bp到100kb。
17.复制模板多核苷酸的方法，该方法包括：体外结合单链模板多核苷酸与来自模板多核苷酸裂解和变性的小随机单链片段，在形成双链模板多核苷酸群体的条件下，在核酸聚合酶存在的条件下保温所说的核酸片段混合物。
18.一种产生适于亲和作用筛选或表型筛选的演示肽或演示抗体文库的方法，该方法包括：
(1)获得包含演示肽或演示抗体和编码所说演示肽或演示抗体的相关多核苷酸的第一群选定的文库成员，并获得所说相关多核苷酸或其拷贝，其中所说的相关多核苷酸包含基本相同序列的区域，以及
(2)在适于PCR扩增的条件下合并且片段化所说的相关多核苷酸或拷贝，进行PCR扩增，且由此同源重组所说的片段使形成重组多核苷酸的杂合库，其中所说的杂合库的重组多核苷酸的主要部分不存在于第一群选定的文库成员中。
19.权利要求18的方法，进一步包含将突变导入所说的多核苷酸或拷贝中。
20.权利要求19的方法，其中经过进行PCR扩增导入突变。
21.权利要求20的方法，其中PCR扩增是有误差的PCR。
22.权利要求18的方法，包含另一步筛选杂合库的文库成员以鉴定具有结合预定大分子之能力的单个杂合文库成员。
23.权利要求18的方法，其中经过选择不同于对预定分子的结合亲和力的表型特征来获得第一群选定的文库成员。
24.权利要求18的方法，其中合并第一群选定的文库成员并片段化，经体外PCR同源重组。
25.权利要求18的方法，其中合并第一群选定的文库成员并体外片段化，所得的片段转移进宿主细胞或有机体并同源重组形成体内杂合文库成员。
26.权利要求18的方法，其中第一群选定的文库成员在游离的可复制载体上克隆或扩增，所说的多重性载体转移进细胞和同源重组以形成体内杂合文库成员。
27.产生适于亲和作用筛选或表型筛选的演示肽或演示抗体的文库的方法，该方法包括：
(1)获得包含演示肽或演示抗体和编码所说演示肽或演示抗体的相关多核苷酸的第一群选定的文库成员，获得所说相关多核苷酸或其拷贝，其中所说的相关多核苷酸包含基本相同序列的区域；且
(2)在游离可复制载体上克隆或扩增所说相关多核苷酸或拷贝并将所说的多重性载体转移进细胞和同源重组以形成体内杂合文库成员。
28.权利要求27的方法，进一步包含将突变导入所说的多核苷酸或其拷贝中。
29.权利要求27的方法，其中所说的游离可复制载体包含许多相关多核苷酸或其拷贝的直接重复。
30.一种产生适于亲和作用筛选的演示抗体的文库的方法，该方法包含：
(1)获得包含演示抗体和编码所说的演示抗体的相关多核苷酸的第一群选定的文库成员，获得所说的相关多核苷酸或其拷贝，其中所说的相关多核苷酸包含基本相同的可变区框架序列的区域；以及
(2)在适于PCR扩增的条件下合并和片段化所说的相关多核苷酸或拷贝以形成其片段，进行PCR扩增，并由此同源重组所说的片段以形成包含新CDR组合的重组多核苷酸的杂合库，其中包含CDR组合的所说杂合库的重组多核苷酸主要部分不存在于第一群选定的文库成员中。
31.权利要求30的方法，包含另一个步骤，其中对杂合库进行亲和筛选以选择结合预定抗原决定基的杂合文库成员，并由此选择许多选定的杂合文库成员。
32.权利要求31的方法，包含另一步杂合许多选定的杂合文库成员并从1到大约1000次循环进行筛选。
CN95191679A 1994-02-17 1995-02-17 通过随机片段化和重装配进行的dna诱变 Pending CN1145641A (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/198,431 1994-02-17
US08/198,431 US5605793A (en) 1994-02-17 1994-02-17 Methods for in vitro recombination

Publications (1)

Publication Number Publication Date
CN1145641A true CN1145641A (zh) 1997-03-19

Family

ID=22733354

Family Applications (1)

Application Number Title Priority Date Filing Date
CN95191679A Pending CN1145641A (zh) 1994-02-17 1995-02-17 通过随机片段化和重装配进行的dna诱变

Country Status (12)

Country Link
US (14) US5605793A (zh)
EP (4) EP0934999B1 (zh)
JP (6) JPH10500561A (zh)
CN (1) CN1145641A (zh)
AT (2) ATE216722T1 (zh)
AU (3) AU703264C (zh)
CA (1) CA2182393C (zh)
DE (5) DE69526497T2 (zh)
DK (2) DK0934999T3 (zh)
ES (2) ES2165652T3 (zh)
RU (1) RU2157851C2 (zh)
WO (1) WO1995022625A1 (zh)

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101258244B (zh) * 2005-08-08 2012-02-15 基因技术股份公司 用于在体内进行蛋白质的选择性进化的方法
CN104805508A (zh) * 2015-04-29 2015-07-29 江南大学 一种基于合成单链dna文库进化代谢途径的方法
CN104837994A (zh) * 2012-12-06 2015-08-12 安捷伦科技有限公司 分子制造
WO2016058127A1 (zh) * 2014-10-14 2016-04-21 深圳华大基因科技有限公司 一种转座酶打断核酸并加接头的方法和试剂
CN105934541A (zh) * 2013-11-27 2016-09-07 Gen9股份有限公司 核酸文库及其制造方法
US10081807B2 (en) 2012-04-24 2018-09-25 Gen9, Inc. Methods for sorting nucleic acids and multiplexed preparative in vitro cloning
US10202608B2 (en) 2006-08-31 2019-02-12 Gen9, Inc. Iterative nucleic acid assembly using activation of vector-encoded traits
US10308931B2 (en) 2012-03-21 2019-06-04 Gen9, Inc. Methods for screening proteins using DNA encoded chemical libraries as templates for enzyme catalysis
CN110366593A (zh) * 2017-02-21 2019-10-22 苏黎世联邦理工学院 Dna零件，途径和基因组的进化指导的多重dna装配
US10457935B2 (en) 2010-11-12 2019-10-29 Gen9, Inc. Protein arrays and methods of using and making the same
US11072789B2 (en) 2012-06-25 2021-07-27 Gen9, Inc. Methods for nucleic acid assembly and high throughput sequencing
US11084014B2 (en) 2010-11-12 2021-08-10 Gen9, Inc. Methods and devices for nucleic acids synthesis
US11702662B2 (en) 2011-08-26 2023-07-18 Gen9, Inc. Compositions and methods for high fidelity assembly of nucleic acids

Families Citing this family (2260)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0834576B1 (en) * 1990-12-06 2002-01-16 Affymetrix, Inc. (a Delaware Corporation) Detection of nucleic acid sequences
US6150141A (en) * 1993-09-10 2000-11-21 Trustees Of Boston University Intron-mediated recombinant techniques and reagents
US20060257890A1 (en) * 1996-05-20 2006-11-16 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US6995017B1 (en) 1994-02-17 2006-02-07 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US5837458A (en) * 1994-02-17 1998-11-17 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US6335160B1 (en) * 1995-02-17 2002-01-01 Maxygen, Inc. Methods and compositions for polypeptide engineering
US6117679A (en) 1994-02-17 2000-09-12 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US5605793A (en) 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination
US6395547B1 (en) 1994-02-17 2002-05-28 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6406855B1 (en) * 1994-02-17 2002-06-18 Maxygen, Inc. Methods and compositions for polypeptide engineering
US6309883B1 (en) * 1994-02-17 2001-10-30 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US6165793A (en) 1996-03-25 2000-12-26 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6808904B2 (en) * 1994-06-16 2004-10-26 Syngenta Participations Ag Herbicide-tolerant protox genes produced by DNA shuffling
DE69535829D1 (de) 1994-08-20 2008-10-16 Gendaq Ltd Verbesserung in bezug auf bindungsproteine bei der erkennung von dna
GB9824544D0 (en) 1998-11-09 1999-01-06 Medical Res Council Screening system
US7285416B2 (en) * 2000-01-24 2007-10-23 Gendaq Limited Regulated gene expression in plants
US7262055B2 (en) 1998-08-25 2007-08-28 Gendaq Limited Regulated gene expression in plants
US6063562A (en) * 1994-09-16 2000-05-16 Sepracor, Inc. In vitro method for predicting the evolutionary response of HIV protease to a drug targeted thereagainst
US7820798B2 (en) * 1994-11-07 2010-10-26 Human Genome Sciences, Inc. Tumor necrosis factor-gamma
US7597886B2 (en) * 1994-11-07 2009-10-06 Human Genome Sciences, Inc. Tumor necrosis factor-gamma
CA2206451C (en) * 1994-12-09 2002-11-26 Wakunaga Seiyaku Kabushiki Kaisha Method for suppressing nonspecific hybridization in primer extension method
US20010053523A1 (en) * 1995-06-02 2001-12-20 M&E Biotech A/S. Method for identification of biologically active peptides and nucleic acids
US6720140B1 (en) * 1995-06-07 2004-04-13 Invitrogen Corporation Recombinational cloning using engineered recombination sites
US6936289B2 (en) 1995-06-07 2005-08-30 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
US6143557A (en) * 1995-06-07 2000-11-07 Life Technologies, Inc. Recombination cloning using engineered recombination sites
US6964861B1 (en) * 1998-11-13 2005-11-15 Invitrogen Corporation Enhanced in vitro recombinational cloning of using ribosomal proteins
US6495357B1 (en) * 1995-07-14 2002-12-17 Novozyme A/S Lipolytic enzymes
US6004788A (en) 1995-07-18 1999-12-21 Diversa Corporation Enzyme kits and libraries
US6455254B1 (en) 1995-07-18 2002-09-24 Diversa Corporation Sequence based screening
US6057103A (en) * 1995-07-18 2000-05-02 Diversa Corporation Screening for novel bioactivities
US5958672A (en) 1995-07-18 1999-09-28 Diversa Corporation Protein activity screening of clones having DNA from uncultivated microorganisms
JP4156666B2 (ja) * 1995-08-11 2008-09-24 ノボザイムス アクティーゼルスカブ ポリペプチド変異体を調整するための方法
DK1143006T3 (da) 1995-08-18 2008-07-14 Morphosys Ip Gmbh Vektorer/DNA-sekvenser fra humane kombinatoriske antistofbiblioteker
US6361974B1 (en) 1995-12-07 2002-03-26 Diversa Corporation Exonuclease-mediated nucleic acid reassembly in directed evolution
US5830696A (en) 1996-12-05 1998-11-03 Diversa Corporation Directed evolution of thermophilic enzymes
US6740506B2 (en) 1995-12-07 2004-05-25 Diversa Corporation End selection in directed evolution
US5965408A (en) 1996-07-09 1999-10-12 Diversa Corporation Method of DNA reassembly by interrupting synthesis
US6790605B1 (en) * 1995-12-07 2004-09-14 Diversa Corporation Methods for obtaining a desired bioactivity or biomolecule using DNA libraries from an environmental source
US6939689B2 (en) 1995-12-07 2005-09-06 Diversa Corporation Exonuclease-mediated nucleic acid reassembly in directed evolution
US20020028443A1 (en) * 1999-09-27 2002-03-07 Jay M. Short Method of dna shuffling with polynucleotides produced by blocking or interrupting a synthesis or amplification process
US6238884B1 (en) 1995-12-07 2001-05-29 Diversa Corporation End selection in directed evolution
US6713279B1 (en) 1995-12-07 2004-03-30 Diversa Corporation Non-stochastic generation of genetic vaccines and enzymes
US20020031771A1 (en) * 1995-12-07 2002-03-14 Short Jay M. Sequence based screening
US20030219752A1 (en) * 1995-12-07 2003-11-27 Diversa Corporation Novel antigen binding molecules for therapeutic, diagnostic, prophylactic, enzymatic, industrial, and agricultural applications, and methods for generating and screening thereof
US6352842B1 (en) 1995-12-07 2002-03-05 Diversa Corporation Exonucease-mediated gene assembly in directed evolution
GB9621295D0 (en) * 1995-12-07 1996-11-27 Cambridge Antibody Tech Specific binding members,materials and methods
US5939250A (en) * 1995-12-07 1999-08-17 Diversa Corporation Production of enzymes having desired activities by mutagenesis
US20020164580A1 (en) * 1995-12-07 2002-11-07 Diversa Corporation Combinatorial screening of mixed populations of organisms
US6489145B1 (en) 1996-07-09 2002-12-03 Diversa Corporation Method of DNA shuffling
US6344328B1 (en) 1995-12-07 2002-02-05 Diversa Corporation Method for screening for enzyme activity
US6358709B1 (en) 1995-12-07 2002-03-19 Diversa Corporation End selection in directed evolution
US6764835B2 (en) * 1995-12-07 2004-07-20 Diversa Corporation Saturation mutageneis in directed evolution
US6537776B1 (en) 1999-06-14 2003-03-25 Diversa Corporation Synthetic ligation reassembly in directed evolution
US7018793B1 (en) 1995-12-07 2006-03-28 Diversa Corporation Combinatorial screening of mixed populations of organisms
US7888466B2 (en) 1996-01-11 2011-02-15 Human Genome Sciences, Inc. Human G-protein chemokine receptor HSATU68
US5976846A (en) * 1996-01-13 1999-11-02 Passmore; Steven E. Method for multifragment in vivo cloning and mutation mapping
WO1997030150A2 (en) * 1996-02-15 1997-08-21 Pangenetics B.V. Molecules for the induction of immunological tolerance
US6096548A (en) 1996-03-25 2000-08-01 Maxygen, Inc. Method for directing evolution of a virus
US6506602B1 (en) 1996-03-25 2003-01-14 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6372480B1 (en) 1996-04-19 2002-04-16 Mycogen Corporation Pesticidal proteins
US6242211B1 (en) 1996-04-24 2001-06-05 Terragen Discovery, Inc. Methods for generating and screening novel metabolic pathways
US7465571B1 (en) 1996-05-22 2008-12-16 Verenium Corporation Endoglucanases
US20020120118A1 (en) * 1996-05-22 2002-08-29 Short Jay M. Enzymes having endoglucanase activity and methods of use thereof
US6617434B1 (en) * 1996-05-31 2003-09-09 North Shore Long Island Jewish Research Institute Identificiaton of differentially methylated and mutated nucleic acids
US5871917A (en) 1996-05-31 1999-02-16 North Shore University Hospital Research Corp. Identification of differentially methylated and mutated nucleic acids
CA2259954C (en) 1996-07-10 2012-03-06 Lonza Ag Process for the preparation of (s)- or (r)-3,3,3-trifluoro-2-hydroxy-2-methylpropionic acid
JPH1066576A (ja) * 1996-08-07 1998-03-10 Novo Nordisk As 突出末端を有する２本鎖ｄｎａ及びこれを用いたｄｎａのシャフリング方法
JPH1066577A (ja) * 1996-08-07 1998-03-10 Novo Nordisk As 核酸プールおよびその製造方法
GB9618960D0 (en) 1996-09-11 1996-10-23 Medical Science Sys Inc Proteases
EP0963434A4 (en) * 1996-09-27 2000-10-25 Maxygen Inc METHODS FOR OPTIMIZING GENE THERAPY THROUGH RECURRENT SEARCHING AND SEQUENCE SELECTION
US5811381A (en) * 1996-10-10 1998-09-22 Mark A. Emalfarb Cellulase compositions and methods of use
US7883872B2 (en) * 1996-10-10 2011-02-08 Dyadic International (Usa), Inc. Construction of highly efficient cellulase compositions for enzymatic hydrolysis of cellulose
US6838238B1 (en) 1996-10-17 2005-01-04 Invitrogen Corporation Morphatides: novel shape and structure libraries
US6242669B1 (en) 1996-10-30 2001-06-05 Mycogen Corporation Pesticidal toxins and nucleotide sequences which encode these toxins
IL119586A (en) * 1996-11-07 2001-09-13 Univ Ramot Discontinuous library of a single biological unit and a method for its preparation
US20070009930A1 (en) * 1996-12-18 2007-01-11 Maxygen, Inc. Methods and compositions for polypeptide engineering
US6326204B1 (en) 1997-01-17 2001-12-04 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
US7148054B2 (en) * 1997-01-17 2006-12-12 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
US6265201B1 (en) 1997-01-17 2001-07-24 Regents Of The University Of Minnesota DNA molecules and protein displaying improved triazine compound degrading ability
IL130635A0 (en) 1997-01-17 2000-06-01 Maxygen Inc Evolution of whole cells and organisms by recursive sequence recombination
AU1022402A (en) * 1997-01-17 2002-03-14 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
US6261804B1 (en) 1997-01-21 2001-07-17 The General Hospital Corporation Selection of proteins using RNA-protein fusions
CN1238366C (zh) 1997-01-21 2006-01-25 综合医院公司 利用rna－蛋白融合体筛选蛋白
US8207093B2 (en) * 1997-01-21 2012-06-26 The General Hospital Corporation Selection of proteins using RNA-protein fusions
GB9701425D0 (en) 1997-01-24 1997-03-12 Bioinvent Int Ab A method for in vitro molecular evolution of protein function
ATE239077T1 (de) * 1997-01-31 2003-05-15 Cosmix Molecular Biolog Gmbh Erzeugung von diversität in kombinatorischen banken
CA2281205A1 (en) 1997-02-12 1998-08-13 Eugene Y. Chan Methods and products for analyzing polymers
WO1998038326A1 (en) 1997-02-28 1998-09-03 Nature Technology Corporation Self-assembling genes, vectors and uses thereof
US5851808A (en) 1997-02-28 1998-12-22 Baylor College Of Medicine Rapid subcloning using site-specific recombination
US6291165B1 (en) * 1997-03-18 2001-09-18 Novo Nordisk A/S Shuffling of heterologous DNA sequences
IL131986A0 (en) * 1997-03-18 2001-03-19 Novo Nordisk As Shuffling of heterologous dna sequences
US6159688A (en) 1997-03-18 2000-12-12 Novo Nordisk A/S Methods of producing polynucleotide variants
US6159687A (en) * 1997-03-18 2000-12-12 Novo Nordisk A/S Methods for generating recombined polynucleotides
AU6611498A (en) * 1997-03-18 1998-10-12 Novo Nordisk A/S Method for constructing a library using dna shuffling
US5948653A (en) * 1997-03-21 1999-09-07 Pati; Sushma Sequence alterations using homologous recombination
US6153410A (en) * 1997-03-25 2000-11-28 California Institute Of Technology Recombination of polynucleotide sequences using random or defined primers
EP0985033A4 (en) * 1997-04-04 2005-07-13 Biosite Inc POLYVALENT AND POLYCLONAL LIBRARIES
CA2286262A1 (en) 1997-04-11 1998-10-22 California Institute Of Technology Apparatus and method for automated protein design
US6558901B1 (en) * 1997-05-02 2003-05-06 Biomerieux Vitek Nucleic acid assays
IL124275A (en) * 1997-05-02 2002-03-10 Bio Merieux Vitek Inc A method to produce sequences of nucleic acids
GB9712512D0 (en) 1997-06-16 1997-08-20 Bioinvent Int Ab A method for in vitro molecular evolution of protein function
US7087420B1 (en) 1997-07-17 2006-08-08 Cambia Microbial β-glucuronidase genes, gene products and uses thereof
US6391547B1 (en) * 1997-09-09 2002-05-21 Center For The Application Of Molecular Biology To International Agriculture Microbial β-glucuronidase genes, gene products and uses thereof
DE19731990A1 (de) * 1997-07-25 1999-01-28 Studiengesellschaft Kohle Mbh Verfahren zur Herstellung und Identifizierung von neuen Hydrolasen mit verbesserten Eigenschaften
WO1999011818A1 (fr) * 1997-08-28 1999-03-11 Isao Karube Procede de detection d'acides nucleiques ou de polypeptides hautement fonctionnels
WO1999014336A2 (en) * 1997-09-19 1999-03-25 Promega Corporation Thermostable luciferases and methods of production
US6602677B1 (en) 1997-09-19 2003-08-05 Promega Corporation Thermostable luciferases and methods of production
ES2322825T3 (es) 1997-10-13 2009-06-29 Novozymes A/S Mutantes de alfa-amilasa.
GB9722131D0 (en) 1997-10-20 1997-12-17 Medical Res Council Method
ATE341621T1 (de) * 1997-10-24 2006-10-15 Invitrogen Corp Rekombinatorisches klonieren unter verwendung von nukleinsaüren mit rekombinationsstellen
US7351578B2 (en) * 1999-12-10 2008-04-01 Invitrogen Corp. Use of multiple recombination sites with unique specificity in recombinational cloning
CN101125873A (zh) * 1997-10-24 2008-02-20 茵维特罗根公司 利用具重组位点的核酸进行重组克隆
US20030124555A1 (en) * 2001-05-21 2003-07-03 Invitrogen Corporation Compositions and methods for use in isolation of nucleic acid molecules
WO1999021979A1 (en) 1997-10-28 1999-05-06 Maxygen, Inc. Human papillomavirus vectors
US20030082110A1 (en) * 1997-10-31 2003-05-01 Vincent Fischetti Use of bacterial phage associated lysing proteins for treating bacterial dental caries
US6406692B1 (en) 1997-10-31 2002-06-18 New Horizons Diagnostics Corp Composition for treatment of an ocular bacterial infection
US6752988B1 (en) * 2000-04-28 2004-06-22 New Horizons Diagnostic Corp Method of treating upper resiratory illnesses
US6428784B1 (en) 1997-10-31 2002-08-06 New Horizons Diagnostics Corp Vaginal suppository for treating group B Streptococcus infection
US20030129146A1 (en) * 1997-10-31 2003-07-10 Vincent Fischetti The use of bacterial phage associated lysing proteins for treating bacterial dental caries
US6432444B1 (en) 1997-10-31 2002-08-13 New Horizons Diagnostics Corp Use of bacterial phage associated lysing enzymes for treating dermatological infections
US20020136712A1 (en) * 1997-10-31 2002-09-26 Fischetti Vincent Bacterial phage associated lysing enzymes for the prophylactic and therapeutic treatment of colonization and infections caused by streptococcus pneumoniae
US6399097B1 (en) 1997-10-31 2002-06-04 New Horizons Diagnostics Corporation Composition for treatment of a bacterial infection of the digestive tract
EP1030861A4 (en) * 1997-10-31 2001-09-05 Maxygen Inc MODIFICATION OF VIRAL TROPISM AND THE DIVERSITY OF HOST SPECIES BY RECOMBINATION OF THE VIRAL GENOME
US20030129147A1 (en) * 1997-10-31 2003-07-10 Vincent Fischetti Use of bacterial phage associated lysing proteins for treating bacterial dental caries
US6277399B1 (en) 1997-10-31 2001-08-21 New Horizon Diagnostics Corporation Composition incorporating bacterial phage associated lysing enzymes for treating dermatological infections
US6056954A (en) 1997-10-31 2000-05-02 New Horizons Diagnostics Corp Use of bacterial phage associated lysing enzymers for the prophylactic and therapeutic treatment of various illnesses
US6423299B1 (en) 1997-10-31 2002-07-23 Vincent Fischetti Composition for treatment of a bacterial infection of an upper respiratory tract
US6399098B1 (en) 1997-10-31 2002-06-04 New Horizons Diagnostics Corp Composition for treating dental caries caused by streptococcus mutans
US7232576B2 (en) 1997-10-31 2007-06-19 New Horizons Diagnostics Corp Throat lozenge for the treatment of Streptococcus Group A
US7321023B2 (en) * 1997-11-07 2008-01-22 Incyte Corporation SP16 protein
JP4139561B2 (ja) * 1997-12-05 2008-08-27 ユーロペーイシェ ラボラトリウム フュール モレキュラーバイオロジー（イーエムビーエル） 新規ｄｎａクローニング方法
JP3712255B2 (ja) 1997-12-08 2005-11-02 カリフォルニア・インスティチュート・オブ・テクノロジー ポリヌクレオチドおよびポリペプチド配列を生成するための方法
AR017831A1 (es) * 1997-12-10 2001-10-24 Pioneer Hi Bred Int Metodo para alterar la composicion de aminoacidos de una proteina nativa de interes, proteina elaborada, y polinucleotido
US20030027173A1 (en) * 1998-01-16 2003-02-06 Della-Cioppa Guy Method of determining the function of nucleotide sequences and the proteins they encode by transfecting the same into a host
US20030166169A1 (en) * 1998-01-16 2003-09-04 Padgett Hal S. Method for constructing viral nucleic acids in a cell-free manner
US6426185B1 (en) * 1998-01-16 2002-07-30 Large Scale Biology Corporation Method of compiling a functional gene profile in a plant by transfecting a nucleic acid sequence of a donor plant into a different host plant in an anti-sense orientation
US6303848B1 (en) 1998-01-16 2001-10-16 Large Scale Biology Corporation Method for conferring herbicide, pest, or disease resistance in plant hosts
US20020164585A1 (en) * 1998-01-16 2002-11-07 Sean Chapman Method for enhancing RNA or protein production using non-native 5' untranslated sequences in recombinant viral nucleic acids
US6468745B1 (en) 1998-01-16 2002-10-22 Large Scale Biology Corporation Method for expressing a library of nucleic acid sequence variants and selecting desired traits
US7390619B1 (en) * 1998-02-11 2008-06-24 Maxygen, Inc. Optimization of immunomodulatory properties of genetic vaccines
US6541011B2 (en) 1998-02-11 2003-04-01 Maxygen, Inc. Antigen library immunization
TR200002537T2 (tr) * 1998-03-17 2000-11-21 Novartis Ag. Mlo proteinlerini kodlayan ve bitki üzerinde mantar direnci veren genler
AU2003203147B2 (en) 1998-03-23 2008-05-29 Novozymes A/S Phytase variants
JP2002513550A (ja) 1998-05-01 2002-05-14 マキシジェン， インコーポレイテッド Ｄｎａシャッフリングを使用する病害虫抵抗性遺伝子の至適化
US6287765B1 (en) * 1998-05-20 2001-09-11 Molecular Machines, Inc. Methods for detecting and identifying single molecules
US6902918B1 (en) 1998-05-21 2005-06-07 California Institute Of Technology Oxygenase enzymes and screening method
US20030207345A1 (en) * 1998-05-21 2003-11-06 California Institute Of Technology Oxygenase enzymes and screening method
EP2287318B1 (en) 1998-06-10 2014-01-22 Novozymes A/S Mannanases
US7153655B2 (en) * 1998-06-16 2006-12-26 Alligator Bioscience Ab Method for in vitro molecular evolution of protein function involving the use of exonuclease enzyme and two populations of parent polynucleotide sequence
EP1090024A2 (en) 1998-06-17 2001-04-11 Maxygen, Inc. Method for producing polynucleotides with desired properties
US6365408B1 (en) 1998-06-19 2002-04-02 Maxygen, Inc. Methods of evolving a polynucleotides by mutagenesis and recombination
US6846655B1 (en) 1998-06-29 2005-01-25 Phylos, Inc. Methods for generating highly diverse libraries
ES2188190T5 (es) 1998-07-21 2007-11-16 Danisco A/S Producto alimentario.
US20030153042A1 (en) * 1998-07-28 2003-08-14 California Institute Of Technology Expression of functional eukaryotic proteins
US20060242731A1 (en) * 1998-08-12 2006-10-26 Venkiteswaran Subramanian DNA shuffling to produce herbicide selective crops
CA2333914A1 (en) * 1998-08-12 2000-02-24 Maxygen, Inc. Dna shuffling to produce herbicide selective crops
US20030104417A1 (en) * 1998-08-12 2003-06-05 Proteus S.A. Template-mediated, ligation-oriented method of nonrandomly shuffling polynucleotides
US20040191772A1 (en) * 1998-08-12 2004-09-30 Dupret Daniel Marc Method of shuffling polynucleotides using templates
US6951719B1 (en) 1999-08-11 2005-10-04 Proteus S.A. Process for obtaining recombined nucleotide sequences in vitro, libraries of sequences and sequences thus obtained
US20030092023A1 (en) * 1998-08-12 2003-05-15 Daniel Dupret Method of shuffling polynucleotides using templates
US6991922B2 (en) * 1998-08-12 2006-01-31 Proteus S.A. Process for in vitro creation of recombinant polynucleotide sequences by oriented ligation
EP1104459A1 (en) 1998-08-12 2001-06-06 Maxygen, Inc. Dna shuffling of monooxygenase genes for production of industrial chemicals
US6602685B1 (en) 1998-08-17 2003-08-05 Phylos, Inc. Identification of compound-protein interactions using libraries of protein-nucleic acid fusion molecules
EP1109931A4 (en) * 1998-09-03 2004-12-15 Univ Loma Linda METHOD FOR STUDYING PROTEIN INTERACTIONS $ i (IN VIVO)
US20070178475A1 (en) * 1998-09-17 2007-08-02 Nehls Michael C Novel human polynucleotides and polypeptides encoded thereby
US20050086718A1 (en) 1999-03-23 2005-04-21 Mendel Biotechnology, Inc. Plant transcriptional regulators of abiotic stress
US7897843B2 (en) 1999-03-23 2011-03-01 Mendel Biotechnology, Inc. Transcriptional regulation of plant biomass and abiotic stress tolerance
ES2237159T3 (es) * 1998-10-06 2005-07-16 Mark Aaron Emalfarb Sistema de transformacion en el campo de los hongos micoticos filamentosos en chrysosporium.
AU6510799A (en) 1998-10-07 2000-04-26 Maxygen, Inc. Dna shuffling to produce nucleic acids for mycotoxin detoxification
US6403312B1 (en) 1998-10-16 2002-06-11 Xencor Protein design automatic for protein libraries
JP2003527072A (ja) * 1998-10-16 2003-09-16 ゼンコー 蛋白質ライブラリーに関する蛋白質設計オートメーション
US20020048772A1 (en) 2000-02-10 2002-04-25 Dahiyat Bassil I. Protein design automation for protein libraries
US7315786B2 (en) 1998-10-16 2008-01-01 Xencor Protein design automation for protein libraries
US7479472B1 (en) 1998-10-19 2009-01-20 The Board Of Trustees Of The Leland Stanford Junior University DNA-templated combinatorial library chemistry
WO2000024883A1 (en) 1998-10-26 2000-05-04 Novozymes A/S Constructing and screening a dna library of interest in filamentous fungal cells
GB9823468D0 (en) 1998-10-28 1998-12-23 Secr Defence Novel enzyme
JP3267576B2 (ja) * 1998-11-09 2002-03-18 三光純薬株式会社 プローブ高分子の形成方法
AU1619400A (en) 1998-11-10 2000-05-29 Maxygen, Inc. Modified adp-glucose pyrophosphorylase for improvement and optimization of plantphenotypes
US6438561B1 (en) * 1998-11-19 2002-08-20 Navigation Technologies Corp. Method and system for using real-time traffic broadcasts with navigation systems
US6303342B1 (en) 1998-11-20 2001-10-16 Kason Biosciences, Inc. Recombinant methods and materials for producing epothilones C and D
US6410301B1 (en) 1998-11-20 2002-06-25 Kosan Biosciences, Inc. Myxococcus host cells for the production of epothilones
ATE338120T2 (de) 1998-11-27 2006-09-15 Ucb Sa Zusammensetzungen und verfahren zur erhöhung der knochenmineralisierung
US20040009535A1 (en) 1998-11-27 2004-01-15 Celltech R&D, Inc. Compositions and methods for increasing bone mineralization
EP1803817B1 (en) 1998-12-18 2011-04-06 Novozymes A/S Subtilase enzymes of the I-S1 and I-S2 sub-groups having an additional amino acid residue in an active site loop region
US20020038013A1 (en) * 2000-02-04 2002-03-28 Gregory Donoho Novel human membrane proteins and polynucleotides encoding the same
US20020031802A1 (en) * 2000-05-12 2002-03-14 Yi Hu Novel seven transmembrane proteins and polynucleotides encoding the same
US6570003B1 (en) * 2001-01-09 2003-05-27 Lexion Genetics Incorporated Human 7TM proteins and polynucleotides encoding the same
US20020061556A1 (en) * 2000-04-27 2002-05-23 Walke D. Wade Novel membrane proteins and polynucleotides encoding the same
US20020055626A1 (en) * 2000-06-09 2002-05-09 Turner C. Alexander Novel human seven transmembrane proteins and polynucleotides encoding the same
US6358712B1 (en) 1999-01-05 2002-03-19 Trustee Of Boston University Ordered gene assembly
US20040005673A1 (en) * 2001-06-29 2004-01-08 Kevin Jarrell System for manipulating nucleic acids
AU774306B2 (en) * 1999-01-05 2004-06-24 Trustees Of Boston University Improved nucleic acid cloning
US7070934B2 (en) 1999-01-12 2006-07-04 Sangamo Biosciences, Inc. Ligand-controlled regulation of endogenous gene expression
US6534261B1 (en) * 1999-01-12 2003-03-18 Sangamo Biosciences, Inc. Regulation of endogenous gene expression in cells using zinc finger proteins
US7013219B2 (en) 1999-01-12 2006-03-14 Sangamo Biosciences, Inc. Regulation of endogenous gene expression in cells using zinc finger proteins
US6436675B1 (en) 1999-09-28 2002-08-20 Maxygen, Inc. Use of codon-varied oligonucleotide synthesis for synthetic shuffling
US6368861B1 (en) * 1999-01-19 2002-04-09 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US20030054390A1 (en) * 1999-01-19 2003-03-20 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
AU2415200A (en) * 1999-01-18 2000-08-01 Maxygen, Inc. Methods of populating data structures for use in evolutionary simulations
US6917882B2 (en) * 1999-01-19 2005-07-12 Maxygen, Inc. Methods for making character strings, polynucleotides and polypeptides having desired characteristics
US6376246B1 (en) 1999-02-05 2002-04-23 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US8457903B1 (en) 1999-01-19 2013-06-04 Codexis Mayflower Holdings, Llc Method and/or apparatus for determining codons
US7702464B1 (en) 2001-08-21 2010-04-20 Maxygen, Inc. Method and apparatus for codon determining
AU3210200A (en) 1999-01-19 2000-08-01 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US6961664B2 (en) 1999-01-19 2005-11-01 Maxygen Methods of populating data structures for use in evolutionary simulations
US7873477B1 (en) * 2001-08-21 2011-01-18 Codexis Mayflower Holdings, Llc Method and system using systematically varied data libraries
US20070065838A1 (en) * 1999-01-19 2007-03-22 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US7024312B1 (en) * 1999-01-19 2006-04-04 Maxygen, Inc. Methods for making character strings, polynucleotides and polypeptides having desired characteristics
US20090130718A1 (en) * 1999-02-04 2009-05-21 Diversa Corporation Gene site saturation mutagenesis
US7384387B1 (en) 1999-02-11 2008-06-10 Maxygen, Inc. High throughput mass spectrometry
CA2363779A1 (en) 1999-02-26 2000-08-31 Human Genome Sciences, Inc. Human endokine alpha and methods of use
AU774643B2 (en) 1999-03-02 2004-07-01 Invitrogen Corporation Compositions and methods for use in recombinational cloning of nucleic acids
JP3399518B2 (ja) * 1999-03-03 2003-04-21 インターナショナル・ビジネス・マシーンズ・コーポレーション 半導体構造およびその製造方法
US6531316B1 (en) 1999-03-05 2003-03-11 Maxyag, Inc. Encryption of traits using split gene sequences and engineered genetic elements
EP1165775A2 (en) * 1999-03-05 2002-01-02 Maxygen, Inc. Recombination of insertion modified nucleic acids
AU3623800A (en) * 1999-03-08 2000-09-28 Protogene Laboratories, Inc. Methods and compositions for economically synthesizing and assembling long dna sequences
IL145165A0 (en) * 1999-03-09 2002-06-30 Diversa Corp End selection in directed evolution
KR100312070B1 (ko) * 1999-03-15 2001-11-03 박호군 고속 선별법을 포함하는 개선된 형질을 갖는 효소 변이체의 제조방법
US8148110B2 (en) * 1999-03-15 2012-04-03 The Board Of Trustees Of The Leland Stanford Junior University Detection of molecular interactions by β-lactamase reporter fragment complementation
US7105315B2 (en) * 1999-06-16 2006-09-12 Incyte Genomics, Inc. Transmembrane protein differentially expressed in cancer
US7030215B2 (en) * 1999-03-24 2006-04-18 Sangamo Biosciences, Inc. Position dependent recognition of GNN nucleotide triplets by zinc fingers
US20030104526A1 (en) 1999-03-24 2003-06-05 Qiang Liu Position dependent recognition of GNN nucleotide triplets by zinc fingers
DK2290060T3 (en) 1999-03-30 2017-03-06 Novozymes As The alpha-amylase variants
AU781258B2 (en) 1999-03-31 2005-05-12 Novozymes A/S Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same
ES2322426T3 (es) 1999-03-31 2009-06-22 Novozymes A/S Polipeptidos con actividad alfa-amilasa y acidos nucleicos que codifican a los mismos.
US6703240B1 (en) 1999-04-13 2004-03-09 Maxygar, Inc. Modified starch metabolism enzymes and encoding genes for improvement and optimization of plant phenotypes
US20020110809A1 (en) * 1999-04-30 2002-08-15 Nehls Michael C. Novel human polynucleotides and polypeptides encoded thereby
US20020095031A1 (en) * 1999-05-04 2002-07-18 Nehls Michael C. Novel human polynucleotides and polypeptides encoded thereby
WO2000071691A1 (en) 1999-05-20 2000-11-30 Novozymes A/S Subtilase enzymes of the i-s1 and i-s2 sub-groups having at least one additional amino acid residue between positions 125 and 126
EP1183342B2 (en) 1999-05-20 2012-06-13 Novozymes A/S Subtilase enzymes of the i-s1 and i-s2 sub-groups having at least one additional amino acid residue between positions 128 and 129
DE60040284D1 (de) 1999-05-20 2008-10-30 Novozymes As Subtilase enzyme der i-s1 und i-s2 untergruppen mit mindestens einer zusätzlichen aminosäure zwischen position 129 und 130
AU4392500A (en) 1999-05-20 2000-12-12 Novozymes A/S Subtilase enzymes of the i-s1 and i-s2 sub-groups having at least one additionalamino acid residue between positions 132 and 133
ATE407201T1 (de) 1999-05-20 2008-09-15 Novozymes As Subtilase-enzyme der i-s1 und i-s2 untergruppen mit wenigstens einem zusätzlichen aminosäurerest zwischen positionen 126 und 127
DE60040285D1 (de) 1999-05-20 2008-10-30 Novozymes As Subtilase enzyme der i-s1 und i-s2 untergruppen mit mindestens einer zusätzlichen aminosäure zwischen position 127 und 128
US7332308B1 (en) 1999-05-21 2008-02-19 The Penn State Research Foundation Incrementally truncated nucleic acids and methods of making same
JP2003509010A (ja) * 1999-05-31 2003-03-11 ノボザイムス アクティーゼルスカブ ペプチドのスクリーニング方法
EP1067181A1 (en) * 1999-06-10 2001-01-10 Rijksuniversiteit te Groningen Enzyme selection
AU1824101A (en) 1999-06-18 2001-01-09 Elitra Pharmaceuticals, Inc. Nucleotide sequences of moraxella catarrhalis genome
WO2001004287A1 (en) * 1999-07-07 2001-01-18 Maxygen Aps A method for preparing modified polypeptides
JP2003504046A (ja) 1999-07-09 2003-02-04 ノボザイムス アクティーゼルスカブ グルコアミラーゼ変異体
US6421613B1 (en) * 1999-07-09 2002-07-16 Pioneer Hi-Bred International, Inc. Data processing of the maize prolifera genetic sequence
US6387620B1 (en) * 1999-07-28 2002-05-14 Gilead Sciences, Inc. Transcription-free selex
US6713309B1 (en) 1999-07-30 2004-03-30 Large Scale Proteomics Corporation Microarrays and their manufacture
US6653151B2 (en) 1999-07-30 2003-11-25 Large Scale Proteomics Corporation Dry deposition of materials for microarrays using matrix displacement
US7179638B2 (en) 1999-07-30 2007-02-20 Large Scale Biology Corporation Microarrays and their manufacture by slicing
GB9918154D0 (en) * 1999-08-02 1999-10-06 Zeneca Ltd Expression system
ATE341632T1 (de) 1999-08-12 2006-10-15 Pfizer Prod Inc Streptomyces avermitilis gen zum regulieren des verhältnisses der b2:b1 avermectinen
US6251604B1 (en) 1999-08-13 2001-06-26 Genopsys, Inc. Random mutagenesis and amplification of nucleic acid
US6720142B1 (en) * 1999-08-19 2004-04-13 University Of Rochester Method of determining evolutionary potential of mutant resistance genes and use thereof to screen for drug efficacy
US6448388B1 (en) 2000-08-16 2002-09-10 Lexicon Genetics Incorporated Human proteases and polynucleotides encoding the same
US20020161214A1 (en) * 2000-11-01 2002-10-31 Walke D. Wade Novel human proteases and polynucleotides encoding the same
US20020049312A1 (en) * 2000-05-23 2002-04-25 Turner C. Alexander Noel human thrombospondin-like proteins and polynucleotides encoding the same
US6716614B1 (en) * 1999-09-02 2004-04-06 Lexicon Genetics Incorporated Human calcium dependent proteases, polynucleotides encoding the same, and uses thereof
US20050153323A1 (en) * 2000-07-28 2005-07-14 Yi Hu Novel human proteases and polynucleotides encoding the same
US20080003673A1 (en) * 1999-09-02 2008-01-03 Alejandro Abuin Novel human proteases and polynucleotides encoding the same
US6649399B2 (en) * 1999-12-09 2003-11-18 Lexicon Genetics Incorporated Human proteases and polynucleotides encoding the same
US20030166902A1 (en) * 2001-02-20 2003-09-04 Yi Hu Novel human protease and polynucleotides encoding the same
US20030050464A1 (en) * 2000-07-28 2003-03-13 Yi Hu Novel human proteases and polynucleotides encoding the same
US6472588B1 (en) * 1999-09-10 2002-10-29 Texas Tech University Transgenic cotton plants with altered fiber characteristics transformed with a sucrose phosphate synthase nucleic acid
US20020127215A1 (en) * 1999-09-14 2002-09-12 Lawrence Loomis Parenteral use of bacterial phage associated lysing enzymes for the therapeutic treatment of bacterial infections
US7063837B2 (en) * 1999-09-14 2006-06-20 New Horizons Diagnostics Corp Syrup composition containing phage associated lytic enzymes
US6867291B1 (en) * 2000-09-15 2005-03-15 Lexicon Genetics Incorporated Human hemicentin proteins and polynucleotides encoding the same
US6790660B1 (en) * 2001-09-18 2004-09-14 Lexicon Genetics Incorporated Human kielin-like proteins and polynucleotides encoding the same
US6579710B2 (en) * 2000-12-12 2003-06-17 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
US6716616B1 (en) 1999-09-28 2004-04-06 Lexicon Genetics Incorporated Human kinase proteins and polynucleotides encoding the same
US20080050809A1 (en) * 1999-09-28 2008-02-28 Alejandro Abuin Novel human kinases and polynucleotides encoding the same
US6586230B1 (en) * 2000-10-27 2003-07-01 Lexicon Genetics Incorporated Human kinase and polynucleotides encoding the same
US6759527B2 (en) 2001-03-20 2004-07-06 Lexicon Genetics Incorporated Human kinase and polynucleotides encoding the same
US6841377B1 (en) 2001-06-13 2005-01-11 Lexicon Genetics Incorporated Human kinase and polynucleotides encoding the same
US6511840B1 (en) 2000-06-15 2003-01-28 Lexicon Genetics Incorporated Human kinase proteins and polynucleotides encoding the same
US6797510B1 (en) * 2001-05-24 2004-09-28 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
US6541252B1 (en) 2000-05-19 2003-04-01 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
US20040002474A1 (en) * 1999-10-07 2004-01-01 Maxygen Inc. IFN-alpha homologues
AU782326B2 (en) 1999-10-12 2005-07-21 Lexicon Pharmaceuticals, Inc. Human LDL receptor family proteins and polynucleotides encoding the same
US7430477B2 (en) * 1999-10-12 2008-09-30 Maxygen, Inc. Methods of populating data structures for use in evolutionary simulations
AU780784B2 (en) 1999-10-13 2005-04-14 Board Of Trustees Of The Leland Stanford Junior University Biosynthesis of polyketide synthase substrates
WO2001027275A1 (en) * 1999-10-13 2001-04-19 Lexicon Genetics Incorporated Novel human membrane proteins
US20080213878A1 (en) * 1999-10-19 2008-09-04 Gregory Donoho Novel human membrane proteins and polynucleotides encoding the same
US6750054B2 (en) 2000-05-18 2004-06-15 Lexicon Genetics Incorporated Human semaphorin homologs and polynucleotides encoding the same
US6346249B1 (en) * 1999-10-22 2002-02-12 Ludwig Institute For Cancer Research Methods for reducing the effects of cancers that express A33 antigen using A33 antigen specific immunoglobulin products
GB9925161D0 (en) 1999-10-26 1999-12-22 Secr Defence Novel enzyme
US6635438B2 (en) * 1999-11-02 2003-10-21 Catch, Inc. Enzymatic cycling assays for homocysteine and cystathionine
EP1230269A2 (en) 1999-11-03 2002-08-14 Maxygen, Inc. Antibody diversity generation
DE19953854C2 (de) 1999-11-09 2002-01-17 Max Planck Gesellschaft Verfahren zur Herstellung von Biopolymeren mit veränderten Eigenschaften
AR026433A1 (es) 1999-11-10 2003-02-12 Novozymes As Variantes de alfa-amilasa tipo fungamyl
US6686515B1 (en) 1999-11-23 2004-02-03 Maxygen, Inc. Homologous recombination in plants
ATE359364T1 (de) * 1999-11-26 2007-05-15 Basf Plant Science Gmbh Verfahren zur mutagenese von nukleotidsequenzen aus pflanzen, algen, oder pilzen
US7115712B1 (en) * 1999-12-02 2006-10-03 Maxygen, Inc. Cytokine polypeptides
JP2004504005A (ja) * 1999-12-07 2004-02-12 レキシコン・ジェネティクス・インコーポレーテッド ヒト新規キナーゼタンパクおよびそれをコードするポリヌクレオチド
WO2001042287A2 (en) * 1999-12-07 2001-06-14 Lexicon Genetics Incorporated Putative human g-protein coupled receptors
DK1250453T3 (da) 1999-12-10 2008-08-11 Invitrogen Corp Anvendelse af multiple rekombinations-sites med unik specificitet i rekombinationskloning
AU2594301A (en) * 1999-12-23 2001-07-03 Maxygen, Inc. Alteration of hydrolase genes and screening of the resulting libraries for the ability to catalyze specific reactions
CA2396029A1 (en) * 1999-12-27 2001-07-05 Crucell Holland B.V. Use of a native epitope for selecting evolved binding members from a library of mutants of a protein capable of binding to said epitope
US6927046B1 (en) 1999-12-30 2005-08-09 Archer-Daniels-Midland Company Increased lysine production by gene amplification using coryneform bacteria
US6395485B1 (en) * 2000-01-11 2002-05-28 Aventis Cropscience N.V. Methods and kits for identifying elite event GAT-ZM1 in biological samples
AU2788101A (en) * 2000-01-11 2001-07-24 Maxygen, Inc. Integrated systems and methods for diversity generation and screening
EP1248846A2 (en) * 2000-01-18 2002-10-16 Lexicon Genetics Incorporated Human kinase protein and polynucleotides encoding the same
JP2003520050A (ja) 2000-01-24 2003-07-02 フィロス インク． タンパク質分析のための高感度多重化診断アッセイ法
US7022479B2 (en) * 2000-01-24 2006-04-04 Compound Therapeutics, Inc. Sensitive, multiplexed diagnostic assays for protein analysis
EP1621617A1 (en) * 2000-02-10 2006-02-01 Xencor, Inc. Protein design automation for protein libraries
WO2001059066A2 (en) * 2000-02-10 2001-08-16 Xencor, Inc. Protein design automation for protein libraries
WO2001064864A2 (en) * 2000-02-28 2001-09-07 Maxygen, Inc. Single-stranded nucleic acid template-mediated recombination and nucleic acid fragment isolation
US20020155571A1 (en) * 2000-02-29 2002-10-24 Raillard Sun Ai Triazine degrading enzymes
US6436677B1 (en) * 2000-03-02 2002-08-20 Promega Corporation Method of reverse transcription
EP1263967A2 (en) 2000-03-10 2002-12-11 Lexicon Genetics Incorporated Human g-coupled protein receptor kinases and polynucleotides encoding the same
AU2001287273A1 (en) * 2000-03-24 2001-10-08 Maxygen, Inc. Methods for modulating cellular and organismal phenotypes
US6767736B2 (en) 2000-04-03 2004-07-27 Lexicon Genetics Incorporated Human ion channel protein and polynucleotides encoding the same
EP1274720A4 (en) 2000-04-12 2004-08-18 Human Genome Sciences Inc ALBUMIN FUSED PROTEINS
JP2003532391A (ja) * 2000-04-12 2003-11-05 レキシコン・ジェネティクス・インコーポレーテッド ヒトメタロプロテアーゼ及び該タンパク質をコードするポリヌクレオチド
US7129326B2 (en) * 2000-04-14 2006-10-31 Genencor International, Inc. Methods for selective targeting
AU2001257012A1 (en) * 2000-04-14 2001-10-30 Genencor International, Inc. Methods for selective targeting
AU2001259205A1 (en) * 2000-04-28 2001-11-12 New Horizons Diagnostic Corporation The use of bacterial phage associated lysing enzymes for treating various illnesses
US6534292B1 (en) 2000-05-08 2003-03-18 Genencor International, Inc. Methods for forming recombined nucleic acids
EP1280909A2 (en) * 2000-05-12 2003-02-05 Lexicon Genetics Incorporated Human lipocalin homologs and polynucleotides encoding the same
US7115403B1 (en) 2000-05-16 2006-10-03 The California Institute Of Technology Directed evolution of galactose oxidase enzymes
US7244560B2 (en) * 2000-05-21 2007-07-17 Invitrogen Corporation Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites
US20030032059A1 (en) * 2000-05-23 2003-02-13 Zhen-Gang Wang Gene recombination and hybrid protein development
US6790667B1 (en) 2000-05-30 2004-09-14 Lexicon Genetics Incorporated Human mitochondrial proteins and polynucleotides encoding the same
US20030031675A1 (en) 2000-06-06 2003-02-13 Mikesell Glen E. B7-related nucleic acids and polypeptides useful for immunomodulation
WO2001094583A2 (en) * 2000-06-07 2001-12-13 Lexicon Genetics Incorporated Human transporter proteins and polynucleotides encoding the same
US6465632B1 (en) * 2000-06-09 2002-10-15 Lexicon Genetics Incorporated Human phosphatases and polynucleotides encoding the same
FR2810339B1 (fr) * 2000-06-14 2004-12-10 Hoechst Marion Roussel Inc Banques combinatoires ameliorees par recombinaison dans la levure et procede d'analyse
EP1294949A4 (en) 2000-06-15 2004-08-25 Human Genome Sciences Inc HUMAN TUMORNESCROSE FACTOR DELTA AND EPSILON
US7468475B2 (en) 2000-06-16 2008-12-23 Schmuelling Thomas Method for modifying plant morphology, biochemistry and physiology
ATE494304T1 (de) 2000-06-16 2011-01-15 Human Genome Sciences Inc Immunspezifisch bindende antikörper gegen blys
US6406863B1 (en) 2000-06-23 2002-06-18 Genetastix Corporation High throughput generation and screening of fully human antibody repertoire in yeast
US6410246B1 (en) 2000-06-23 2002-06-25 Genetastix Corporation Highly diverse library of yeast expression vectors
US6410271B1 (en) 2000-06-23 2002-06-25 Genetastix Corporation Generation of highly diverse library of expression vectors via homologous recombination in yeast
AU2001268716A1 (en) * 2000-06-23 2002-01-08 Maxygen, Inc. Novel chimeric promoters
WO2002000717A2 (en) * 2000-06-23 2002-01-03 Maxygen, Inc. Novel co-stimulatory molecules
WO2002000852A2 (en) 2000-06-26 2002-01-03 Novozymes A/S Lipolytic enzyme
US7846733B2 (en) 2000-06-26 2010-12-07 Nugen Technologies, Inc. Methods and compositions for transcription-based nucleic acid amplification
CA2415761A1 (en) * 2000-06-27 2002-01-03 Lexicon Genetics Incorporated Novel human gaba receptors and polynucleotides encoding the same
WO2002004629A2 (en) * 2000-07-07 2002-01-17 Maxygen, Inc. Molecular breeding of transposable elements
US6994963B1 (en) 2000-07-10 2006-02-07 Ambion, Inc. Methods for recombinatorial nucleic acid synthesis
US6858422B2 (en) * 2000-07-13 2005-02-22 Codexis, Inc. Lipase genes
WO2002006469A2 (en) * 2000-07-18 2002-01-24 Enchira Biotechnology Corporation Methods of ligation mediated chimeragenesis utilizing populations of scaffold and donor nucleic acids
US6878861B2 (en) * 2000-07-21 2005-04-12 Washington State University Research Foundation Acyl coenzyme A thioesterases
US7435562B2 (en) 2000-07-21 2008-10-14 Modular Genetics, Inc. Modular vector systems
AU2002224553A1 (en) * 2000-07-21 2002-02-05 Incyte Genomics, Inc. Human proteases
US6887685B1 (en) 2000-07-25 2005-05-03 Lexicon Genetics Incorporated Human thymosin protein and polynucleotides encoding the same
AU2001279135A1 (en) * 2000-07-31 2002-02-13 Maxygen, Inc. Biosensors, reagents and diagnostic applications of directed evolution
AU2001280968A1 (en) * 2000-07-31 2002-02-13 Menzel, Rolf Compositions and methods for directed gene assembly
JP4855632B2 (ja) 2000-08-01 2012-01-18 ノボザイムス アクティーゼルスカブ 変更された性質を有するα−アミラーゼ突然変異体
JP3859947B2 (ja) * 2000-08-04 2006-12-20 独立行政法人理化学研究所 突然変異導入方法
AU8337701A (en) 2000-08-14 2002-02-25 Us Health Enhanced homologous recombination mediated by lambda recombination proteins
AR030462A1 (es) 2000-08-21 2003-08-20 Novozymes As Novedosas enzimas subtilasas que tienen una reducida tendencia hacia su inhibicion por las sustancias presentes en los huevos
US7198924B2 (en) * 2000-12-11 2007-04-03 Invitrogen Corporation Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites
JP2004506447A (ja) * 2000-08-22 2004-03-04 レキシコン・ジェネティクス・インコーポレーテッド 新規ヒト７ｔｍタンパクおよびそれをコードするポリヌクレオチド
EP1406483A4 (en) 2000-08-22 2005-05-25 Mendel Biotechnology Inc GENE FOR CHANGING THE PROPERTIES OF PLANTS
EP1311690B1 (en) * 2000-08-22 2006-07-26 Lexicon Genetics Incorporated Human proteases and polynucleotides encoding the same
US20030157643A1 (en) * 2000-08-24 2003-08-21 Almond Brian D Synthetic nucleic acids from aquatic species
WO2002016583A2 (en) * 2000-08-24 2002-02-28 Maxygen, Inc. Constructs and their use in metabolic pathway engineering
US7879540B1 (en) * 2000-08-24 2011-02-01 Promega Corporation Synthetic nucleic acid molecule compositions and methods of preparation
FR2813314B1 (fr) * 2000-08-25 2004-05-07 Biomethodes Procede de mutagenese dirigee massive
IL154596A0 (en) 2000-08-25 2003-09-17 Basf Plant Science Gmbh Plant polynucleotides encoding novel prenyl proteases
AU2001285326A1 (en) * 2000-08-31 2002-03-13 Lexicon Genetics Incorporated Human kinase proteins and polynucleotides encoding the same
US6395504B1 (en) 2000-09-01 2002-05-28 New Horizons Diagnostics Corp. Use of phage associated lytic enzymes for the rapid detection of bacterial contaminants
US7229790B2 (en) * 2000-09-01 2007-06-12 Lexicon Pharmaceuticals, Inc. Human GABA transporter protein and polynucleotides encoding the same
AU2001290744A1 (en) * 2000-09-11 2002-03-26 University Of Rochester Method of identifying putative antibiotic resistance genes
WO2002024953A1 (en) * 2000-09-21 2002-03-28 Merck & Co., Inc. Method for generating recombinant polynucleotides
US20020142324A1 (en) * 2000-09-22 2002-10-03 Xun Wang Fungal target genes and methods to identify those genes
US7820441B2 (en) * 2000-09-25 2010-10-26 The Regents Of The University Of Michigan Production of viral vectors
AU2001293084A1 (en) * 2000-09-27 2002-04-08 Lexicon Genetics Incorporated Human protease inhibitor proteins and polynucleotides encoding the same
JPWO2002026964A1 (ja) * 2000-09-27 2004-02-05 三菱化学株式会社 変異ｄｎａライブラリーの構築方法およびその利用
US20040091920A1 (en) * 2000-09-27 2004-05-13 Mitsubishi Chemical Corporation Method of constructing a mutant DNA library and use thereof
AU2001293019B2 (en) * 2000-09-27 2007-01-18 Lexicon Pharmaceuticals, Inc. Human ion-exchanger proteins and polynucleotides encoding the same
US6777232B1 (en) 2000-10-02 2004-08-17 Lexicon Genetics Incorporated Human membrane proteins and polynucleotides encoding the same
WO2002033089A2 (en) * 2000-10-05 2002-04-25 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Ixodes scapularis tissue factor pathway inhibitor
WO2002031745A1 (en) * 2000-10-10 2002-04-18 Genencor International, Inc. Information rich libraries
US7122310B2 (en) * 2000-10-11 2006-10-17 Sanko Junyaku Co., Ltd. Methods of constructing self-assembly of probes and method of detecting the same
WO2002031129A2 (en) 2000-10-12 2002-04-18 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
JP4198987B2 (ja) 2000-10-13 2008-12-17 ノボザイムス アクティーゼルスカブ サブチラーゼ変異体
US20040091994A1 (en) 2000-10-13 2004-05-13 Carsten Andersen Alpha-amylase variant with altered properties
US7001763B1 (en) 2000-10-17 2006-02-21 Lexicon Genetics Incorporated Human semaphorin proteins and polynucleotides encoding the same
US7045283B2 (en) 2000-10-18 2006-05-16 The Regents Of The University Of California Methods of high-throughput screening for internalizing antibodies
WO2002070705A2 (en) * 2000-10-27 2002-09-12 Lexicon Genetics Incorporated Human 7tm proteins and polynucleotides encoding them
WO2002036633A2 (en) * 2000-10-30 2002-05-10 Lexicon Genetics Incorporated Human 7tm proteins and polynucleotides encoding the same
US7060442B2 (en) * 2000-10-30 2006-06-13 Regents Of The University Of Michigan Modulators on Nod2 signaling
AU2004260931B9 (en) 2003-04-29 2012-01-19 E.I. Du Pont De Nemours And Company Novel glyphosate-N-acetyltransferase (GAT) genes
US7202070B2 (en) * 2000-10-31 2007-04-10 Biocatalytics, Inc. Method for reductive amination of a ketone using a mutated enzyme
CA2427340C (en) 2000-10-31 2013-09-10 Zycos Inc. Cyp1b1 nucleic acids and methods of use
WO2002102405A1 (en) * 2000-11-02 2002-12-27 New Horizons Diagnostics Corporation The use of bacterial phage associated lytic enzymes to prevent food poisoning
WO2002038757A1 (en) * 2000-11-10 2002-05-16 Amicogen, Inc. Method for generating recombinant dna library using unidirectional single-stranded dna fragments
JP2004535154A (ja) * 2000-11-15 2004-11-25 レキシコン・ジェネティクス・インコーポレーテッド 新規ヒト分泌タンパク質およびそれをコードするポリヌクレオチド
EP1830190A3 (en) * 2000-11-17 2008-07-09 University Of Rochester In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells
CA2429544C (en) * 2000-11-17 2010-10-19 University Of Rochester In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells
AU2002228633A1 (en) * 2000-11-20 2002-06-03 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
TWI327599B (en) 2000-11-28 2010-07-21 Medimmune Llc Methods of administering/dosing anti-rsv antibodies for prophylaxis and treatment
WO2002044425A2 (en) 2000-12-01 2002-06-06 Visigen Biotechnologies, Inc. Enzymatic nucleic acid synthesis: compositions and methods for altering monomer incorporation fidelity
CA2431007A1 (en) * 2000-12-11 2002-07-18 Lexicon Genetics Incorporated Novel human kinase and polynucleotides encoding the same
US6958213B2 (en) 2000-12-12 2005-10-25 Alligator Bioscience Ab Method for in vitro molecular evolution of protein function
WO2002048402A2 (en) 2000-12-13 2002-06-20 Nugen Technologies, Inc. Methods and compositions for generation of multiple copies of nucleic acid sequences and methods of detection thereof
DE10062086A1 (de) * 2000-12-13 2002-07-04 Wella Ag Mittel und Verfahren zum Färben von Keratinfasern
US6583269B1 (en) 2000-12-18 2003-06-24 Lexicon Genetics Incorporated Human protease inhibitor and polynucleotides encoding the same
US6852844B1 (en) 2000-12-20 2005-02-08 Lexicon Genetics Incorporated Human protocadherin proteins and polynucleotides encoding the same
CA2432737A1 (en) * 2000-12-20 2002-06-27 Lexicon Genetics Incorporated Human ion channel protein and polynucleotides encoding the same
US20020086292A1 (en) 2000-12-22 2002-07-04 Shigeaki Harayama Synthesis of hybrid polynucleotide molecules using single-stranded polynucleotide molecules
US20020150895A1 (en) * 2000-12-22 2002-10-17 Raymond Wheeler Method and apparatus for filtering and extending RNA alignment coverage
US7070973B2 (en) * 2000-12-26 2006-07-04 Board Of Regents Of The University Of Nebraska Butyrylcholinesterase variants and methods of use
AU2002246858A1 (en) * 2000-12-27 2002-08-06 Lexicon Genetics Incorporated Human kinases and polynucleotides encoding the same
WO2002068626A2 (en) * 2000-12-28 2002-09-06 Lexicon Genetics Incorporated Novel human ion channel-related proteins and polynucleotides encoding the same
WO2002053753A2 (en) * 2001-01-05 2002-07-11 Lexicon Genetics Incorporated Novel human lipase and polynucleotides encoding the same
DK1353941T3 (da) * 2001-01-22 2013-06-17 Sangamo Biosciences Inc Modificerede zinkfingerbindingsproteiner
US20030104402A1 (en) * 2001-01-23 2003-06-05 University Of Rochester Methods of producing or identifying intrabodies in eukaryotic cells
WO2002059287A2 (en) * 2001-01-23 2002-08-01 Lexicon Genetics Incorporated Novel human kinases and polynucleotides encoding the same
US20020115846A1 (en) * 2001-01-24 2002-08-22 Xuanchuan (Sean) Yu Novel human lipase and polynucleotides encoding the same
EP1683865A3 (en) 2001-02-02 2006-10-25 Eli Lilly & Company Mammalian proteins and in particular CD200
WO2002062822A2 (en) * 2001-02-02 2002-08-15 University Of Rochester Methods of identifying regulator molecules
US20020127674A1 (en) * 2001-02-02 2002-09-12 Xuanchuan Yu Novel human transporter protein and polynucleotides encoding the same
WO2002064612A2 (en) 2001-02-09 2002-08-22 Human Genome Sciences, Inc. Human g-protein chemokine receptor (ccr5) hdgnr10
US6849449B2 (en) * 2001-03-01 2005-02-01 Regents Of The University Of Michigan Orphanin FQ receptor nucleic acids
ZA200210369B (en) 2001-03-09 2004-07-08 Nugen Technologies Inc Methods and compositions for amplification or RNA sequences.
JP2004535781A (ja) * 2001-03-12 2004-12-02 レキシコン・ジェネティクス・インコーポレーテッド 新規ヒトｅｆｇファミリータンパクおよびそれをコードするポリヌクレオチド
WO2002072774A2 (en) * 2001-03-12 2002-09-19 Lexicon Genetics Incorporated Novel human transporter proteins and polynucleotides encoding the same
WO2002072603A2 (en) * 2001-03-12 2002-09-19 Lexicon Genetics Incorporated Novel human dectin proteins and polynucleotides encoding the same
US6994995B1 (en) 2001-03-16 2006-02-07 Lexicon Genetics Incorporated Human synaptotagmin and polynucleotides encoding the same
US7807408B2 (en) * 2001-03-19 2010-10-05 President & Fellows Of Harvard College Directed evolution of proteins
CA2441820C (en) * 2001-03-19 2014-01-07 President And Fellows Of Harvard College Evolving new molecular function
AU2002257076A1 (en) 2001-03-19 2002-10-03 President And Fellows Of Harvard College Nucleic acid shuffling
US20030049619A1 (en) * 2001-03-21 2003-03-13 Simon Delagrave Methods for the synthesis of polynucleotides and combinatorial libraries of polynucleotides
AR035799A1 (es) 2001-03-30 2004-07-14 Syngenta Participations Ag Toxinas insecticidas aisladas de bacillus thuringiensis y sus usos.
WO2002081671A1 (en) 2001-04-06 2002-10-17 Lexicon Genetics Incorporated Novel human kinase and polynucleotides encoding the same
JP2004524043A (ja) 2001-04-06 2004-08-12 レキシコン・ジェネティクス・インコーポレーテッド 新規ヒトキナーゼ及び該キナーゼをコードするポリヌクレオチド
FR2823219B1 (fr) * 2001-04-10 2003-07-04 Pasteur Institut Mutants de la desoxycytidine kinase possedant une activite enzymatique elargie
US6644173B2 (en) * 2001-04-11 2003-11-11 Keuring, Incorporated Beverage filter cartridge holder
CA2444632A1 (en) 2001-04-13 2002-10-24 Human Genome Sciences, Inc. Vascular endothelial growth factor 2
WO2002083868A2 (en) * 2001-04-16 2002-10-24 California Institute Of Technology Peroxide-driven cytochrome p450 oxygenase variants
US6607895B2 (en) 2001-04-16 2003-08-19 Lexicon Genetics Incorporated Human adenylsuccinate synthetase and polynucleotides encoding the same
US20030077804A1 (en) * 2001-04-19 2003-04-24 Invitrogen Corporation Compositions and methods for recombinational cloning of nucleic acid molecules
FR2824073B1 (fr) * 2001-04-25 2003-12-19 Proteus Procede de creation in vitro de sequences polynucleotidiques recombinees par ligation orientee
EP1381680A1 (en) * 2001-04-25 2004-01-21 Proteus Template-mediated, ligation-oriented method of nonrandomly shuffling polynucleotides
WO2002088313A2 (en) * 2001-04-30 2002-11-07 Lexicon Genetics Incorporated Novel human nuclear transporters and polynucleotides encoding the same
AU2002254773B2 (en) * 2001-05-03 2005-12-08 Rensselaer Polytechnic Institute Novel methods of directed evolution
EP1950305A1 (en) 2001-05-09 2008-07-30 Monsanto Technology, LLC Tyr a genes and uses thereof
WO2002090517A2 (en) * 2001-05-09 2002-11-14 Lexicon Genetics Incorporated Novel human kinases and polynucleotides encoding the same
EP2159279A3 (en) 2001-05-15 2010-05-12 Novozymes A/S Alpha-amylase variant with altered properties
EP1404830B1 (en) * 2001-05-17 2008-07-16 Proteus Method of preparing polynucleotide fragments for use in shuffling
AU2002339115B2 (en) 2001-05-18 2007-03-15 Dupont Nutrition Biosciences Aps Method of preparing a dough with an enzyme
AU2002309989A1 (en) 2001-05-18 2002-12-03 Rigel Pharmaceuticals, Incorporated Directed evolution of protein in mammalian cells
US20020193585A1 (en) * 2001-05-25 2002-12-19 Walke D. Wade Novel human transporter proteins and polynucleotides encoding the same
JP4309758B2 (ja) 2001-05-25 2009-08-05 ヒューマン ジノーム サイエンシーズ， インコーポレイテッド Ｔｒａｉｌレセプターに免疫特異的に結合する抗体
CA2448347C (en) 2001-05-29 2010-09-21 Lexicon Genetics Incorporated Novel human hydroxylases and polynucleotides encoding the same
AU2002314997A1 (en) * 2001-06-05 2002-12-16 Gorilla Genomics, Inc. Methods for low background cloning of dna using long oligonucleotides
WO2002099091A2 (en) 2001-06-06 2002-12-12 Novozymes A/S Endo-beta-1,4-glucanase from bacillus
EP1406645A4 (en) * 2001-06-14 2005-02-16 Lexicon Genetics Inc NOVEL HUMAN TRANSPORTER PROTEINS AND POLYNUCLEOTIDES ENCODING THESE PROTEINS
US7727713B2 (en) 2001-06-20 2010-06-01 Nuevolution A/S Templated molecules and methods for using such molecules
EP1925672A1 (en) 2001-06-22 2008-05-28 Syngeta Participations AG Abiotic stress responsive polynucleotides and polypeptides
BR0210593A (pt) 2001-06-22 2007-01-02 Pioneer Hi Bred Int polinucleotìdeos de defensina e métodos de uso
AU2002316809A1 (en) 2001-06-26 2003-01-08 Novozymes A/S Polypeptides having cellobiohydrolase i activity and polynucleotides encoding same
JP3967319B2 (ja) * 2001-06-27 2007-08-29 エフ．ホフマン−ラ ロシュ アーゲー ポリヌクレオチド配列のインビトロ組換えのためのウォークスルー技術
GB0115841D0 (en) 2001-06-28 2001-08-22 Medical Res Council Ligand
AU2002318186A1 (en) * 2001-07-03 2003-01-21 Lexicon Genetics Incorporated Novel human kielin-like proteins and polynucleotides encoding the same
DK200101090A (da) 2001-07-12 2001-08-16 Novozymes As Subtilase variants
US20040213765A1 (en) * 2001-07-13 2004-10-28 Vincent Fischetti Use of bacterial phage associated lytic enzymes to prevent food poisoning
US7112423B2 (en) * 2001-07-15 2006-09-26 Keck Graduate Institute Nucleic acid amplification using nicking agents
EP1277835A1 (en) * 2001-07-19 2003-01-22 Libragen Methods of creating genetic diversity
WO2003008563A2 (en) 2001-07-20 2003-01-30 California Institute Of Technology Improved cytochrome p450 oxygenases
ATE286124T1 (de) 2001-07-23 2005-01-15 Dsm Ip Assets Bv Verfahren zur herstellung von polynukleotidvarianten
US8067556B2 (en) * 2001-07-26 2011-11-29 Agilent Technologies, Inc. Multi-site mutagenesis
US6867189B2 (en) * 2001-07-26 2005-03-15 Genset S.A. Use of adipsin/complement factor D in the treatment of metabolic related disorders
US6833441B2 (en) 2001-08-01 2004-12-21 Abmaxis, Inc. Compositions and methods for generating chimeric heteromultimers
WO2003012126A2 (en) * 2001-08-03 2003-02-13 Diversa Corporation Epoxide hydrolases, nucleic acids encoding them and methods for making and using them
US6943001B2 (en) * 2001-08-03 2005-09-13 Diversa Corporation Epoxide hydrolases, nucleic acids encoding them and methods for making and using them
WO2003016481A2 (en) * 2001-08-14 2003-02-27 Lexicon Genetics Incorporated Novel human collagen proteins and polynucleotides encoding the same
US6987079B2 (en) 2001-08-14 2006-01-17 W.R. Grace & Co.-Conn. Supported catalyst systems
US20030096275A1 (en) * 2001-08-20 2003-05-22 Laing Lance G. Biosensor for small molecule analytes
US7647184B2 (en) * 2001-08-27 2010-01-12 Hanall Pharmaceuticals, Co. Ltd High throughput directed evolution by rational mutagenesis
WO2003020892A2 (en) 2001-08-29 2003-03-13 Genentech, Inc. Bv8 NUCLEIC ACIDS AND POLYPEPTIDES WITH MITOGENIC ACTIVITY
US7635798B2 (en) * 2001-08-31 2009-12-22 Dow Agrosciences, Llc Nucleic acid compositions conferring altered metabolic characteristics
KR100428998B1 (ko) * 2001-09-10 2004-04-28 한국과학기술원 모체 단백질과는 크기와 서열이 다른 단백질의 변이집단을생산하는 방법
CA2460546A1 (en) * 2001-09-14 2003-03-27 Invitrogen Corporation Dna polymerases and mutants thereof
EP1438428A4 (en) * 2001-09-14 2005-11-16 Univ Queensland EVIDENCE OF METHYLATION OF DNA
US7108975B2 (en) * 2001-09-21 2006-09-19 Regents Of The University Of Michigan Atlastin
US7582425B2 (en) * 2001-09-21 2009-09-01 The Regents Of The University Of Michigan Atlastin
US20030104445A1 (en) * 2001-09-25 2003-06-05 Hayes Robert J. RNA dependent RNA polymerase mediated protein evolution
DE10147074A1 (de) * 2001-09-25 2003-05-08 Beru Ag Verfahren zum Betreiben einer aus mehreren Heizelementen bestehenden mehrstufigen elektrischen Heizung
WO2003029456A1 (en) 2001-10-01 2003-04-10 Dyax Corp. Multi-chain eukaryotic display vectors and uses thereof
US7157277B2 (en) 2001-11-28 2007-01-02 Neose Technologies, Inc. Factor VIII remodeling and glycoconjugation of Factor VIII
ES2556338T3 (es) 2001-10-10 2016-01-15 Novo Nordisk A/S Remodelación y glicoconjugación de péptidos
US7214660B2 (en) 2001-10-10 2007-05-08 Neose Technologies, Inc. Erythropoietin: remodeling and glycoconjugation of erythropoietin
US7173003B2 (en) * 2001-10-10 2007-02-06 Neose Technologies, Inc. Granulocyte colony stimulating factor: remodeling and glycoconjugation of G-CSF
EP2305311A3 (en) 2001-10-10 2011-07-20 BioGeneriX AG Glycoconjugation of peptides
US8008252B2 (en) 2001-10-10 2011-08-30 Novo Nordisk A/S Factor VII: remodeling and glycoconjugation of Factor VII
AU2004236174B2 (en) 2001-10-10 2011-06-02 Novo Nordisk A/S Glycopegylation methods and proteins/peptides produced by the methods
US20040005709A1 (en) * 2001-10-24 2004-01-08 Hoogenboom Henricus Renerus Jacobus Mattheus Hybridization control of sequence variation
DK1440083T3 (da) 2001-10-25 2013-03-25 Medical Res Council Molekyler
EP1840211A1 (en) 2001-10-31 2007-10-03 Danisco A/S Pyranosone dehydratase from phanerochaete chrysosporium
US7175983B2 (en) * 2001-11-02 2007-02-13 Abmaxis, Inc. Adapter-directed display systems
US6768476B2 (en) * 2001-12-05 2004-07-27 Etenna Corporation Capacitively-loaded bent-wire monopole on an artificial magnetic conductor
US6989261B2 (en) * 2001-12-20 2006-01-24 Eli Lilly And Company Butyrylcholinesterase variant polypeptides with increased catalytic efficiency and methods of use
US7049121B2 (en) * 2001-12-20 2006-05-23 Applied Molecular Evolution Butyrylcholinesterase variant polypeptides with increased catalytic efficiency and methods of use
TWI262083B (en) * 2001-12-28 2006-09-21 Syngenta Participations Ag Microbially-expressed thermotolerant phytase for animal feed
KR100475305B1 (ko) * 2002-01-08 2005-03-10 주식회사 마이크로아이디 단일가닥 특이적인 ｄｎａ 절단효소 엑소뉴클리아제ⅶ을 이용하여 키메라 ｄｎａ 라이브러리를 만드는 방법
US20040009498A1 (en) * 2002-01-14 2004-01-15 Diversa Corporation Chimeric antigen binding molecules and methods for making and using them
US20070166704A1 (en) * 2002-01-18 2007-07-19 Fei Huang Identification of polynucleotides and polypeptide for predicting activity of compounds that interact with protein tyrosine kinases and/or protein tyrosine kinase pathways
US7262054B2 (en) * 2002-01-22 2007-08-28 Sangamo Biosciences, Inc. Zinc finger proteins for DNA binding and gene regulation in plants
DE20200926U1 (de) * 2002-01-23 2002-04-18 Hegenscheidt Mfd Gmbh & Co Kg Festwalzgerät einer Festwalzmaschine für Kurbelwellen
US20040096826A1 (en) * 2002-01-30 2004-05-20 Evans Glen A. Methods for creating recombination products between nucleotide sequences
US7255986B2 (en) * 2002-01-31 2007-08-14 The Board Of Trustees Operating Michigan State University Compositions for the diagnosis and treatment of epizootic catarrhal enteritis in ferrets
ATE426659T1 (de) 2002-02-12 2009-04-15 Pfizer Prod Inc Streptomyces avermitilis gen zur regulierung des verhaltnisses der b2:b1 avermectine
MXPA04007886A (es) 2002-02-13 2004-10-15 Dow Global Technologies Inc Sobre-expresion de genes de extremozimo en pseudomonadas y bacterias estrechamente relacionadas.
US20030224404A1 (en) * 2002-02-25 2003-12-04 Manuel Vega High throughput directed evolution of nucleic acids by rational mutagenesis
US20030166010A1 (en) * 2002-02-25 2003-09-04 Affholter Joseph A. Custom ligand design for biomolecular filtration and purification for bioseperation
DE60323607D1 (de) * 2002-02-26 2008-10-30 Du Pont Verfahren zur rekombination genetischer elemente
WO2003072742A2 (en) * 2002-02-27 2003-09-04 California Institute Of Technology Novel glucose 6-oxidases
US6977162B2 (en) 2002-03-01 2005-12-20 Ravgen, Inc. Rapid analysis of variations in a genome
US20040132101A1 (en) 2002-09-27 2004-07-08 Xencor Optimized Fc variants and methods for their generation
US7783428B2 (en) 2002-03-01 2010-08-24 Maxygen, Inc. Methods, systems, and software for identifying functional biomolecules
CA2477611A1 (en) * 2002-03-01 2003-09-12 Ravgen, Inc. Rapid analysis of variations in a genome
US7244592B2 (en) 2002-03-07 2007-07-17 Dyax Corp. Ligand screening and discovery
WO2003078583A2 (en) 2002-03-09 2003-09-25 Maxygen, Inc. Optimization of crossover points for directed evolution
US7176025B2 (en) * 2002-03-11 2007-02-13 Nugen Technologies, Inc. Methods for generating double stranded DNA comprising a 3′ single stranded portion and uses of these complexes for recombination
AU2003253069A1 (en) * 2002-03-15 2003-09-29 Nuevolution A/S A building block forming a c-c bond upon reaction
EP1487849A2 (en) * 2002-03-15 2004-12-22 Nuevolution A/S A building block capable of transferring a functional entity
EP1487850A2 (en) * 2002-03-15 2004-12-22 Nuevolution A/S A building block forming a c-c or a c-hetero atom bond upon reaction
DK1487978T3 (da) 2002-03-15 2009-03-23 Nuevolution As Forbedret fremgangsmåde til syntese af templatemolekyler
US20050034188A1 (en) * 2002-03-20 2005-02-10 J. R. Simplot Company Refined plant transformation
NZ536037A (en) * 2002-03-20 2008-04-30 Simplot Co J R Refined plant transformation
CA2911801A1 (en) 2002-03-22 2003-10-02 Greta Arnaut Novel bacillus thuringiensis insecticidal proteins
US20030180781A1 (en) * 2002-03-25 2003-09-25 The Regents Of The University Of California Constructing very long DNA sequences from synthetic DNA molecules
US20030186301A1 (en) * 2002-03-25 2003-10-02 The Regents Of The University Of California Constructing user-defined, long DNA sequences
US7452666B2 (en) * 2002-03-25 2008-11-18 Lawrence Livermore National Security, Llc Synthesis of DNA
US20030228598A1 (en) * 2002-03-25 2003-12-11 The Regents Of The University Of California DNA/RNA as a write/read medium
EP1497445A2 (en) * 2002-04-01 2005-01-19 Human Genome Sciences, Inc. Antibodies that specifically bind to gmad
WO2003087763A2 (en) 2002-04-03 2003-10-23 Celltech R & D, Inc. Association of polymorphisms in the sost gene region with bone mineral density
EP2270049A3 (en) 2002-04-12 2011-03-09 Medimmune, Inc. Recombinant anti-interleukin-9-antibody
US7226771B2 (en) 2002-04-19 2007-06-05 Diversa Corporation Phospholipases, nucleic acids encoding them and methods for making and using them
CA2481411C (en) 2002-04-19 2016-06-14 Diversa Corporation Phospholipases, nucleic acids encoding them and methods for making and using them
US6776751B2 (en) * 2002-04-22 2004-08-17 Kendor Laboratory Products, Lp Rotor cover attachment apparatus
US20070178478A1 (en) * 2002-05-08 2007-08-02 Dhallan Ravinder S Methods for detection of genetic disorders
US7442506B2 (en) 2002-05-08 2008-10-28 Ravgen, Inc. Methods for detection of genetic disorders
US7727720B2 (en) 2002-05-08 2010-06-01 Ravgen, Inc. Methods for detection of genetic disorders
EP2243837A1 (en) 2002-05-14 2010-10-27 Martek Biosciences Corporation Carotene synthase gene and uses therefor
ES2285118T5 (es) * 2002-05-17 2012-09-21 Alligator Bioscience Ab Un método para el desarrollo molecular in vitro de una función proteica.
DE10222858A1 (de) 2002-05-23 2003-12-04 Basf Ag Verfahren zur fermentativen Herstellung schwefelhaltiger Feinchemikalien
DE60319259T2 (de) * 2002-06-14 2009-03-05 Dyax Corp., Cambridge Rekombination von nukleinsäurebibliotheksmitgliedern
US7132100B2 (en) 2002-06-14 2006-11-07 Medimmune, Inc. Stabilized liquid anti-RSV antibody formulations
US7425618B2 (en) 2002-06-14 2008-09-16 Medimmune, Inc. Stabilized anti-respiratory syncytial virus (RSV) antibody formulations
CA2490004A1 (en) * 2002-06-20 2003-12-31 Board Of Trustees Operating Michigan State University Plastid division and related genes and proteins, and methods of use
DE60336555D1 (de) * 2002-06-21 2011-05-12 Novo Nordisk Healthcare Ag Pegylierte glykoformen von faktor vii
JP2005536198A (ja) 2002-06-28 2005-12-02 ダウ アグロサイエンス リミテッド ライアビリティー カンパニー ペニバチルス種から得られる殺虫活性タンパク質およびポリヌクレオチド
US20060002935A1 (en) 2002-06-28 2006-01-05 Domantis Limited Tumor Necrosis Factor Receptor 1 antagonists and methods of use therefor
US9321832B2 (en) 2002-06-28 2016-04-26 Domantis Limited Ligand
CN1678634A (zh) * 2002-06-28 2005-10-05 多曼蒂斯有限公司 免疫球蛋白单个变体抗原结合区及其特异性构建体
WO2004009761A2 (en) 2002-07-18 2004-01-29 Monsanto Technology Llc Methods for using artificial polynucleotides and compositions thereof to reduce transgene silencing
EP1543128A4 (en) * 2002-07-18 2008-02-20 Invitrogen Corp VIRUS VECTORS WITH RECOMBINATION SITES
EP2264155A1 (en) 2002-07-25 2010-12-22 Bio-Rad Laboratories, Inc. Hybrid polymerase methods and compositions
US7560260B2 (en) * 2002-07-25 2009-07-14 Bio-Rad Laboratories, Inc. Compositions with polymerase activity
AU2003247266A1 (en) 2002-08-01 2004-02-23 Nuevolution A/S Multi-step synthesis of templated molecules
EP1576151A4 (en) 2002-08-06 2006-05-17 Verdia Inc VARIANTS OF AMINE OXIDASE AP1
US20040067532A1 (en) 2002-08-12 2004-04-08 Genetastix Corporation High throughput generation and affinity maturation of humanized antibody
WO2004016750A2 (en) 2002-08-14 2004-02-26 Macrogenics, Inc. FcϜRIIB-SPECIFIC ANTIBODIES AND METHODS OF USE THEREOF
AU2003263937B2 (en) * 2002-08-19 2010-04-01 The President And Fellows Of Harvard College Evolving new molecular function
US7361635B2 (en) 2002-08-29 2008-04-22 Sangamo Biosciences, Inc. Simultaneous modulation of multiple genes
US20050202438A1 (en) * 2002-09-09 2005-09-15 Rene Gantier Rational directed protein evolution using two-dimensional rational mutagenesis scanning
US20060020396A1 (en) * 2002-09-09 2006-01-26 Rene Gantier Rational directed protein evolution using two-dimensional rational mutagenesis scanning
ES2343518T3 (es) * 2002-09-09 2010-08-03 Hanall Biopharma Co., Ltd. Polipeptidos interferon alfa modificados resistentes a proteasas.
US7563600B2 (en) 2002-09-12 2009-07-21 Combimatrix Corporation Microarray synthesis and assembly of gene-length polynucleotides
ES2380017T3 (es) 2002-09-18 2012-05-07 Mendel Biotechnology, Inc. Polinucleótidos y polipéptidos en plantas
EP2364996B1 (en) 2002-09-27 2016-11-09 Xencor Inc. Optimized FC variants and methods for their generation
GB0222846D0 (en) 2002-10-03 2002-11-06 Choo Yen Cell culture
AU2002951899A0 (en) * 2002-10-04 2002-10-24 Macquarie University Random drift mutagenesis
US6863731B2 (en) * 2002-10-18 2005-03-08 Controls Corporation Of America System for deposition of inert barrier coating to increase corrosion resistance
JP3447009B1 (ja) * 2002-10-29 2003-09-16 實 平垣 構築物用構成体およびその製造方法
DK2348124T3 (en) 2002-10-30 2014-03-10 Nuevolution As Synthesis of a bifunctional complex
EP1558745A4 (en) * 2002-11-01 2006-08-30 Evogenix Pty Ltd MUTAGENESIS METHODS USING RIBAVIRIN AND / OR REPLICASED RNA
TWI319007B (en) 2002-11-06 2010-01-01 Novozymes As Subtilase variants
US7365186B2 (en) * 2002-11-22 2008-04-29 Arborgen, Llc Vascular-preferred promoter sequences and uses thereof
CN102584951A (zh) 2002-11-25 2012-07-18 金克克国际有限公司 皮肤或毛发结合肽
CA2507626A1 (en) * 2002-12-04 2004-06-17 Applied Molecular Evolution Inc. Butyrylcholinesterase variants that alter the activity of chemotherapeutic agents
WO2004056994A2 (en) 2002-12-19 2004-07-08 Nuevolution A/S Quasirandom structure and function guided synthesis methods
US7537921B2 (en) 2002-12-20 2009-05-26 Novozymes A/S Galactanase variants
DK1578964T4 (da) 2002-12-20 2013-12-02 Novozymes As Polypeptider med cellobiohydrolase II-aktivitet og polynukleotider, der koder for samme
US20060014248A1 (en) * 2003-01-06 2006-01-19 Xencor, Inc. TNF super family members with altered immunogenicity
US20050221443A1 (en) * 2003-01-06 2005-10-06 Xencor, Inc. Tumor necrosis factor super family agonists
US7553930B2 (en) * 2003-01-06 2009-06-30 Xencor, Inc. BAFF variants and methods thereof
US20050130892A1 (en) * 2003-03-07 2005-06-16 Xencor, Inc. BAFF variants and methods thereof
US7355008B2 (en) 2003-01-09 2008-04-08 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
US20050196766A1 (en) 2003-12-24 2005-09-08 Soe Jorn B. Proteins
MXPA05007653A (es) 2003-01-17 2005-09-30 Danisco Metodo.
CN102021205A (zh) 2003-01-17 2011-04-20 丹尼斯科公司 制备蛋白质酯和蛋白质亚基酯的方法
US7955814B2 (en) 2003-01-17 2011-06-07 Danisco A/S Method
US8709435B2 (en) 2003-01-21 2014-04-29 Biomay Ag Hypallergenic mosaic antigens and methods of making same
AU2004206250B8 (en) * 2003-01-21 2009-09-17 Bristol-Myers Squibb Company Polynucleotide encoding a novel acyl coenzyme a, monoacylglycerol acyltransferase-3 (MGAT3), and uses thereof
NZ567340A (en) 2003-02-20 2008-10-31 Athenix Corp Delta-endotoxin genes of bacillus thuringiensis and methods for their use as pesticide
US20060269920A1 (en) * 2003-02-21 2006-11-30 Nuevolution A/S Method for obtaining structural information about an encoded molecule
US20070026397A1 (en) 2003-02-21 2007-02-01 Nuevolution A/S Method for producing second-generation library
CN104388449A (zh) 2003-03-06 2015-03-04 维莱尼姆公司 淀粉酶、编码它们的核酸及其制备和应用方法
CA2889013C (en) 2003-03-07 2018-07-17 Dsm Ip Assets B.V. Hydrolases, nucleic acids encoding them and methods for making and using them
AU2004219592C1 (en) 2003-03-12 2011-02-24 Genentech, Inc. Use of Bv8 and/or EG-VEGF to promote hematopoiesis
CA2519092C (en) * 2003-03-14 2014-08-05 Neose Technologies, Inc. Branched water-soluble polymers and their conjugates
DK1608748T3 (da) * 2003-03-20 2009-06-29 Nuevolution As Ligeringskodning af små molekyler
US8980646B2 (en) * 2003-03-26 2015-03-17 Sudhir Paul Proteolytic and covalent antibodies
US8017323B2 (en) * 2003-03-26 2011-09-13 President And Fellows Of Harvard College Free reactant use in nucleic acid-templated synthesis
EP1610808A4 (en) 2003-03-26 2011-04-06 Sudhir Paul COVALENT BINDING OF LIGANDS TO NUCLEOPHILIC PROTEINS UNDER GUIDANCE BY NONCOVALENT BINDING
WO2004087882A2 (en) * 2003-03-26 2004-10-14 Washington University Neuregulin protein regulation of synaptic proteins
JP2007524603A (ja) * 2003-03-27 2007-08-30 ザ ユニバーシティー オブ テキサス Ｈｉｖ／エイズの受動免疫治療用ループス抗体
MXPA05010681A (es) 2003-04-04 2005-12-15 Diversa Corp Liasas de pectato, acidos nucleicos que las codifican, y metodos para hacerlas y usarlas.
US8791070B2 (en) 2003-04-09 2014-07-29 Novo Nordisk A/S Glycopegylated factor IX
KR20110094361A (ko) 2003-04-11 2011-08-23 메디뮨 엘엘씨 재조합 ｉｌ­９ 항체 및 그의 용도
WO2004092418A2 (en) 2003-04-14 2004-10-28 Nugen Technologies, Inc. Global amplification using a randomly primed composite primer
US20040214194A1 (en) * 2003-04-17 2004-10-28 Mj Bioworks, Inc. Methods of making hybrid proteins
JP4850698B2 (ja) 2003-04-25 2012-01-11 ノース・キャロライナ・ステイト・ユニヴァーシティ 細胞表面タンパク質相同体をコードするアシドフィルス菌核酸配列及びその使用
US7834147B2 (en) 2003-04-28 2010-11-16 Childrens Hospital Medical Center Saposin C-DOPS: a novel anti-tumor agent
EP1622921B1 (en) 2003-05-02 2010-06-16 Novozymes Inc. Variants of beta-glucosidases
US20050009772A1 (en) * 2003-05-06 2005-01-13 The Regents Of The University Of California Methods and compositions for the treatment of glaucoma and other retinal diseases
CN102766545B (zh) 2003-05-07 2014-08-06 诺维信公司 变体枯草杆菌蛋白酶（枯草杆菌酶）
US7521242B2 (en) 2003-05-09 2009-04-21 The United States Of America As Represented By The Department Of Health And Human Services Host cells deficient for mismatch repair and their use in methods for inducing homologous recombination using single-stranded nucleic acids
ES2380093T3 (es) * 2003-05-09 2012-05-08 Biogenerix Ag Composiciones y métodos para la preparación de mutantes de glucosilación de la hormona del crecimiento humana
EP1628680B1 (en) 2003-05-15 2012-10-24 Novartis Vaccines and Diagnostics, Inc. Hiv polynucleotides and polypeptides derived from botswana mj4
US7173010B2 (en) * 2003-05-19 2007-02-06 Regents Of The University Of Michigan Orphanin FQ receptor
US7655833B2 (en) * 2003-05-29 2010-02-02 Brookhaven Science Associates, Llc ADS genes for reducing saturated fatty acid levels in seed oils
JP4909072B2 (ja) * 2003-06-06 2012-04-04 アーバージェン インコーポレイテッド 転写因子
AU2004252481B2 (en) * 2003-06-06 2011-01-27 Arborgen Inc. Compositions and methods for regulating polysaccharides of a plant cell
US7381409B2 (en) * 2003-06-16 2008-06-03 Celltech R&D, Inc. Antibodies specific for sclerostin and methods of screening and use therefor
WO2005017106A2 (en) * 2003-06-17 2005-02-24 California Institute Of Technology Libraries of optimized cytochrome p450 enzymes and the optimized p450 enzymes
EP1639091B1 (en) 2003-06-17 2012-12-05 California University Of Technology Regio- and enantioselective alkane hydroxylation with modified cytochrome p450
CA2529659A1 (en) 2003-06-18 2004-12-29 Ulrich Haupts Engineered proteolytic enzyme with altered specificity determining regions
DK1639106T3 (da) 2003-06-19 2010-09-27 Novozymes As Proteaser
EP1651750B1 (en) 2003-06-23 2011-04-06 North Carolina State University Lactobacillus acidophilus nucleic acids encoding fructo-oligosaccharide utilization compounds and uses thereof
EP1644538A4 (en) * 2003-06-26 2006-11-08 Invitrogen Corp METHODS AND COMPOSITIONS FOR DETECTING PROMOTER ACTIVITY AND EXPRESSING FUSION PROTEINS
CN108486086A (zh) 2003-07-02 2018-09-04 维莱尼姆公司 葡聚糖酶，编码它们的核酸以及制备和使用它们的方法
WO2005012484A2 (en) 2003-07-25 2005-02-10 Neose Technologies, Inc. Antibody-toxin conjugates
WO2005017116A2 (en) * 2003-08-11 2005-02-24 California Institute Of Technology Thermostable peroxide-driven cytochrome p450 oxygenase variants and methods of use
WO2005021714A2 (en) 2003-08-11 2005-03-10 Diversa Corporation Laccases, nucleic acids encoding them and methods for making and using them
ES2458636T3 (es) 2003-08-18 2014-05-06 Medimmune, Llc Humanización de anticuerpos
US20060228350A1 (en) * 2003-08-18 2006-10-12 Medimmune, Inc. Framework-shuffling of antibodies
US7326782B2 (en) * 2003-08-21 2008-02-05 Wisconsin Alumni Research Foundation Metabolic engineering of viomycin biosynthesis
DK2617826T3 (en) 2003-08-25 2015-10-12 Novozymes Inc Variants of glucosidhydrolase
JP4839215B2 (ja) * 2003-08-25 2011-12-21 ファンザイム バイオテクノロジーズ エスアー 新規真菌タンパク質および同タンパク質をコードする核酸
DK1664343T3 (da) 2003-09-09 2014-08-11 Integrigen Inc Fremgangsmåder og sammensætninger til generering af kimcelle humane antistofgener
WO2005026387A1 (en) 2003-09-18 2005-03-24 Nuevolution A/S A method for obtaining structural information concerning an encoded molecule and method for selecting compounds
EP1689430B1 (en) * 2003-10-07 2009-07-29 United Therapeutics Corporation Tyrosinase mutant and methods of use thereof
WO2005035747A1 (en) 2003-10-10 2005-04-21 Novozymes A/S Protease variants
EP1675871A2 (en) 2003-10-10 2006-07-05 Xencor Inc. Protein based tnf-alpha variants for the treatment of tnf-alpha related disorders
US20050084868A1 (en) * 2003-10-16 2005-04-21 Wolfgang Aehle Generation of stabilized proteins by combinatorial consensus mutagenesis
EP1678296B1 (en) 2003-10-23 2011-07-13 Novozymes A/S Protease with improved stability in detergents
EP1682656B1 (en) 2003-10-28 2013-09-18 Novozymes Inc. Polypeptides having beta-glucosidase activity and polynucleotides encoding same
JP4916886B2 (ja) 2003-11-12 2012-04-18 イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニー 油性植物および酵母中の多価不飽和脂肪酸レベルを変更するのに適したδ１５デサチュラーゼ
US7985569B2 (en) 2003-11-19 2011-07-26 Danisco Us Inc. Cellulomonas 69B4 serine protease variants
CA2546451A1 (en) 2003-11-19 2005-06-09 Genencor International, Inc. Serine proteases, nucleic acids encoding serine enzymes and vectors and host cells incorporating same
AU2003283943A1 (en) * 2003-11-20 2005-06-08 Agency For Science Technology And Research Method
US7842661B2 (en) * 2003-11-24 2010-11-30 Novo Nordisk A/S Glycopegylated erythropoietin formulations
US20080305992A1 (en) 2003-11-24 2008-12-11 Neose Technologies, Inc. Glycopegylated erythropoietin
ES2445948T3 (es) * 2003-11-24 2014-03-06 Ratiopharm Gmbh Eritropoyetina glicopegilada
US8633157B2 (en) * 2003-11-24 2014-01-21 Novo Nordisk A/S Glycopegylated erythropoietin
EP1697534B1 (en) * 2003-12-01 2010-06-02 Life Technologies Corporation Nucleic acid molecules containing recombination sites and methods of using the same
US7956032B2 (en) 2003-12-03 2011-06-07 Novo Nordisk A/S Glycopegylated granulocyte colony stimulating factor
US20060040856A1 (en) * 2003-12-03 2006-02-23 Neose Technologies, Inc. Glycopegylated factor IX
NZ547554A (en) * 2003-12-03 2009-09-25 Biogenerix Ag Glycopegylated granulocyte colony stimulating factor
ATE352616T1 (de) * 2003-12-04 2007-02-15 Hoffmann La Roche Verfahren zur herstellung von zirkulären mutierten und/oder chimären polynukleotiden
US20070169227A1 (en) 2003-12-16 2007-07-19 Pioneer Hi-Bred International Inc. Dominant Gene Suppression Transgenes and Methods of Using Same
GB0716126D0 (en) 2007-08-17 2007-09-26 Danisco Process
US7718408B2 (en) 2003-12-24 2010-05-18 Danisco A/S Method
WO2008090395A1 (en) 2007-01-25 2008-07-31 Danisco A/S Production of a lipid acyltransferase from transformed bacillus licheniformis cells
KR101169265B1 (ko) 2003-12-24 2012-08-02 대니스코 에이/에스 단백질
CN101072789B (zh) * 2004-01-08 2013-05-15 生物种属学股份公司 肽的o-连接的糖基化
US7432057B2 (en) * 2004-01-30 2008-10-07 Michigan State University Genetic test for PSE-susceptible turkeys
DK2305703T3 (da) 2004-01-30 2014-06-16 Novozymes Inc Polypeptider med cellulolytisk forøgende aktivitet og polynukleotider, der koder for disse
EP2322630B1 (en) 2004-02-12 2016-11-16 Novozymes Inc. Polypeptides having xylanase activity and polynucleotides encoding same
EP2184360B1 (en) 2004-02-25 2015-08-26 Pioneer Hi-Bred International Inc. Novel bacillus thuringiensis crystal polypeptides, polynucleotides, and compositions thereof
EP1718742B1 (en) 2004-02-25 2014-12-24 Novozymes A/S Fungal cell wall degrading enzyme
JP2007534320A (ja) * 2004-02-27 2007-11-29 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ ポリヌクレオチド合成法
US7459289B2 (en) 2004-03-08 2008-12-02 North Carolina State University Lactobacillus acidophilus nucleic acid sequences encoding carbohydrate utilization-related proteins and uses therefor
GB0405637D0 (en) 2004-03-12 2004-04-21 Danisco Protein
EP1730277B1 (en) * 2004-03-22 2009-10-28 Nuevolution A/S Ligational encoding using building block oligonucleotides
CA2560613C (en) 2004-03-22 2015-11-24 Solvay Pharmaceuticals Gmbh Oral pharmaceutical compositions of lipase-containing products, in particular of pancreatin, containing surfactants
US7569223B2 (en) * 2004-03-22 2009-08-04 The Rockefeller University Phage-associated lytic enzymes for treatment of Streptococcus pneumoniae and related conditions
EP1786463A4 (en) * 2004-03-26 2009-05-20 Human Genome Sciences Inc ANTIBODY AGAINST NOGO RECEPTOR
EP1732650A4 (en) * 2004-03-27 2008-06-11 Univ Arizona COMPOSITION AND METHOD FOR CANCER TREATMENT
US7790956B2 (en) 2004-04-02 2010-09-07 Cropdesign N.V. Plants having improved growth characteristics and methods for making the same by modulating expression of a nucleic acid sequence encoding a NAP1-like protein
WO2005115477A2 (en) 2004-04-13 2005-12-08 Quintessence Biosciences, Inc. Non-natural ribonuclease conjugates as cytotoxic agents
US7838733B2 (en) 2004-04-30 2010-11-23 Dow Agrosciences Llc Herbicide resistance genes
US7622125B2 (en) 2004-05-05 2009-11-24 Novartis Vaccines And Diagnostics, Inc. Polycistronic HIV vector constructs
KR101083595B1 (ko) * 2004-05-14 2011-11-16 가톨릭대학교 산학협력단 단백질 형질도입 도메인 유전자를 함유하는 재조합베큘로바이러스 발현벡터
US7674621B2 (en) 2004-05-21 2010-03-09 The United States Of America As Represented By The Department Of Health And Human Services Plasmids and phages for homologous recombination and methods of use
DK1766040T3 (da) 2004-05-27 2017-11-13 Novozymes Inc Fremgangsmåder til transformering og ekspressions-screening af filamentøse svampeceller med et DNA-bibliotek
US7825293B2 (en) 2004-05-28 2010-11-02 Cropdesign N.V. Plants having improved growth characteristics and a method for making the same
CA2735315A1 (en) 2004-06-09 2005-12-29 Pioneer Hi-Bred International, Inc. Plastid transit peptides
ES2540728T3 (es) 2004-06-16 2015-07-13 Dsm Ip Assets B.V. Método para la decoloración enzimática de feofitina
US20060008844A1 (en) 2004-06-17 2006-01-12 Avidia Research Institute c-Met kinase binding proteins
EP2258839B1 (en) 2004-06-21 2015-06-03 Novozymes A/S Proteases
US7985892B1 (en) 2004-06-29 2011-07-26 Dow Agrosciences Llc Truncated Cry35 proteins
EP2221366B1 (en) 2004-06-29 2015-06-10 Novozymes Inc. Polypeptides having alpha-glucosidase activity and polynucleotides encoding same
AU2005327983B2 (en) 2004-06-30 2008-08-07 Pioneer Hi-Bred International, Inc. Methods of protecting plants from pathogenic fungi
EP2113512B1 (en) 2004-07-02 2013-04-17 Pioneer Hi-Bred International Inc. Antifungal polypeptides
JP4955546B2 (ja) 2004-07-05 2012-06-20 ノボザイムス アクティーゼルスカブ 変更された性質を有するα−アミラーゼ変異体
US7928288B2 (en) 2004-07-12 2011-04-19 Cropdesign N.V. Plants having improved growth characteristics and method for making the same
WO2006010143A2 (en) 2004-07-13 2006-01-26 Neose Technologies, Inc. Branched peg remodeling and glycosylation of glucagon-like peptide-1 [glp-1]
EP2267108A1 (en) 2004-07-16 2010-12-29 Danisco A/S Enzymatic oil-degumming method
US20090053249A1 (en) 2004-07-20 2009-02-26 Yan Qi Specific Inhibition of Autoimmunity and Diseases Associated With Autoantigens
US20090292110A1 (en) * 2004-07-23 2009-11-26 Defrees Shawn Enzymatic modification of glycopeptides
CA2575878A1 (en) 2004-08-02 2006-02-09 Novozymes A/S Creation of diversity in polypeptides
EP1791975A4 (en) * 2004-08-05 2007-11-07 Biosite Inc COMPOSITIONS AND METHODS FOR EXPRESSION AT THE SURFACE OF POLYPEPTIDE PHAGES
US7597884B2 (en) 2004-08-09 2009-10-06 Alios Biopharma, Inc. Hyperglycosylated polypeptide variants and methods of use
US7148051B2 (en) * 2004-08-16 2006-12-12 E. I. Du Pont De Nemours And Company Production of 3-hydroxycarboxylic acid using nitrilase
FI20050753A (fi) 2004-09-03 2006-03-04 Licentia Oy Uudet peptidit
EP1799249A2 (en) * 2004-09-10 2007-06-27 Neose Technologies, Inc. Glycopegylated interferon alpha
US7563443B2 (en) * 2004-09-17 2009-07-21 Domantis Limited Monovalent anti-CD40L antibody polypeptides and compositions thereof
US7728118B2 (en) 2004-09-17 2010-06-01 Promega Corporation Synthetic nucleic acid molecule compositions and methods of preparation
US7700720B2 (en) 2004-09-21 2010-04-20 Medimmune, Llc Antibodies against and methods for producing vaccines for respiratory syncytial virus
US7453025B2 (en) * 2004-09-22 2008-11-18 Arborgen, Llc Reproductive ablation constructs
US7799906B1 (en) 2004-09-22 2010-09-21 Arborgen, Llc Compositions and methods for modulating lignin of a plant
GB0421598D0 (en) 2004-09-29 2004-10-27 Cambridge Advanced Tech Modification of plant development and morphology
EP1799819B1 (en) 2004-09-30 2011-03-23 Novozymes Inc. Polypeptides having lipase activity and polynucleotides encoding same
WO2006049777A2 (en) * 2004-09-30 2006-05-11 Trustees Of Dartmouth College Nucleic acid sequences encoding luciferase for expression in filamentous fungi
GB0422052D0 (en) 2004-10-04 2004-11-03 Dansico As Enzymes
CA2601422C (en) 2004-10-04 2015-12-29 Novozymes A/S Polypeptides having phytase activity and polynucleotides encoding same
AR050895A1 (es) 2004-10-04 2006-11-29 Novozymes As Polipeptidos que tienen actividad de fitasa y polinucleotidos que los codifican
EP2163632A1 (en) 2004-10-05 2010-03-17 SunGene GmbH Constitutive expression cassettes for regulation of plant expression
BRPI0516556B1 (pt) 2004-10-08 2019-12-31 Dow Agrosciences Llc composição alimentícia frita, processo de obtenção de uma composição alimentícia frita, processo de redução da gordura saturada na fração de óleo de sementes de uma planta transgênica de canola e polinucleotídeo
RU2401842C2 (ru) 2004-10-08 2010-10-20 Домантис Лимитед Антагонисты и способы их применения
GB0423139D0 (en) 2004-10-18 2004-11-17 Danisco Enzymes
US20060141626A1 (en) * 2004-10-27 2006-06-29 Monsanto Technology Llc Non-random method of gene shuffling
AU2005299355A1 (en) 2004-10-27 2006-05-04 Medimmune, Llc Modulation of antibody specificity by tailoring the affinity to cognate antigens
WO2006047791A2 (en) * 2004-10-27 2006-05-04 The Board Of Trustees Of The Leland Stanford Junior University Dna-templated combinatorial library device and method for use
EP1807521B1 (en) 2004-10-29 2010-12-29 CropDesign N.V. Plants having improved growth characteristics and method for making the same
EP1814573B1 (en) * 2004-10-29 2016-03-09 ratiopharm GmbH Remodeling and glycopegylation of fibroblast growth factor (fgf)
EP1655364A3 (en) 2004-11-05 2006-08-02 BASF Plant Science GmbH Expression cassettes for seed-preferential expression in plants
WO2006053131A1 (en) 2004-11-11 2006-05-18 Modular Genetics, Inc. Ladder assembly and system for generating diversity
EP1814996A2 (en) 2004-11-19 2007-08-08 Novozymes A/S Polypeptides having antimicrobial activity and polynucleotides encoding same
EP1662000B1 (en) 2004-11-25 2011-03-30 SunGene GmbH Expression cassettes for guard cell-preferential expression in plants
EP1666599A3 (en) 2004-12-04 2006-07-12 SunGene GmbH Expression cassettes for mesophyll- and/or epidermis-preferential expression in plants
EP1669455B1 (en) 2004-12-08 2009-10-28 SunGene GmbH Expression cassettes for vascular tissue-preferential expression in plants
EP1669456A3 (en) 2004-12-11 2006-07-12 SunGene GmbH Expression cassettes for meristem-preferential expression in plants
US7198927B2 (en) 2004-12-22 2007-04-03 E. I. Du Pont De Nemours And Company Enzymatic production of glycolic acid
DK1831385T3 (en) 2004-12-22 2015-07-13 Novozymes North America Inc Enzymes for starch processing
EP1831376A1 (en) 2004-12-28 2007-09-12 Pioneer Hi-Bred International, Inc. Improved grain quality through altered expression of seed proteins
US20100009902A1 (en) * 2005-01-06 2010-01-14 Neose Technologies, Inc. Glycoconjugation Using Saccharyl Fragments
NZ556436A (en) 2005-01-10 2010-11-26 Biogenerix Ag Glycopegylated granulocyte colony stimulating factor
CA2594728C (en) 2005-01-13 2013-03-19 Pioneer Hi-Bred International, Inc. Maize cyclo1 gene and promoter
US20060160138A1 (en) * 2005-01-13 2006-07-20 George Church Compositions and methods for protein design
CN101111600A (zh) 2005-01-27 2008-01-23 克罗普迪塞恩股份有限公司 产率增加的植物及其制备方法
WO2006089133A2 (en) 2005-02-15 2006-08-24 Duke University Anti-cd19 antibodies and uses in oncology
EP1856266A4 (en) 2005-03-02 2015-04-22 Inst Nac De Tecnologia Agropecuaria HERBICIDE-RESISTANT RICE PLANTS, POLYNUCLEOTIDES ENCODING HIGH HERBICIDE-RESISTANT ACETHYDROXYACID SYNTHASE SUBUNIT PROTEINS, AND METHODS OF USE
CA2614769A1 (en) 2005-03-10 2006-09-21 Verenium Corporation Lyase enzymes, nucleic acids encoding them and methods for making and using them
BRPI0609140A2 (pt) 2005-03-15 2010-02-17 Verenium Corp celulases, ácidos nucléicos que codificam eles e métodos para a produção e uso deles
CN100577805C (zh) * 2005-03-16 2010-01-06 中国科学院沈阳应用生态研究所 一种基因改组的方法及所获得的重组基因和编码蛋白
JP5153613B2 (ja) 2005-03-18 2013-02-27 メディミューン，エルエルシー 抗体のフレームワーク・シャッフル
AU2006224944B2 (en) 2005-03-18 2012-01-12 Novozymes Adenium Biotech A/S Polypeptides having antimicrobial activity and polynucleotides encoding same
CA2601400A1 (en) 2005-03-19 2006-09-28 Medical Research Council Improvements in or relating to treatment and prevention of viral infections
US11214817B2 (en) 2005-03-28 2022-01-04 California Institute Of Technology Alkane oxidation by modified hydroxylases
US8715988B2 (en) 2005-03-28 2014-05-06 California Institute Of Technology Alkane oxidation by modified hydroxylases
AU2006232196B2 (en) 2005-04-01 2012-11-22 Athenix Corporation AXMI-027, AXMI-036 and AXMI-038, a family of delta-endotoxin genes and methods for their use
US9187546B2 (en) * 2005-04-08 2015-11-17 Novo Nordisk A/S Compositions and methods for the preparation of protease resistant human growth hormone glycosylation mutants
EP3479844B1 (en) 2005-04-15 2023-11-22 MacroGenics, Inc. Covalent diabodies and uses thereof
BRPI0610609A2 (pt) 2005-04-15 2010-07-06 Univ North Carolina State métodos e composições para modular a adesão e toleráncia ao estresse em bactérias
CA2606220A1 (en) 2005-04-19 2006-12-21 Basf Plant Science Gmbh Starchy-endosperm and/or germinating embryo-specific expression in mono-cotyledonous plants
CA2606475C (en) 2005-04-27 2015-06-16 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
US7592429B2 (en) 2005-05-03 2009-09-22 Ucb Sa Sclerostin-binding antibody
US8003108B2 (en) 2005-05-03 2011-08-23 Amgen Inc. Sclerostin epitopes
EP2221316A1 (en) 2005-05-05 2010-08-25 Duke University Anti-CD19 antibody therapy for autoimmune disease
CN1861789B (zh) * 2005-05-13 2010-04-28 中国科学院沈阳应用生态研究所 一种综合的基因改组方法及所获得的重组基因和编码蛋白
AR053269A1 (es) 2005-05-16 2007-04-25 Monsanto Technology Llc Plantas y semillas de maiz con mejoramiento de asparagina y proteina
US20060281113A1 (en) * 2005-05-18 2006-12-14 George Church Accessible polynucleotide libraries and methods of use thereof
US20060263974A1 (en) * 2005-05-18 2006-11-23 Micron Technology, Inc. Methods of electrically interconnecting different elevation conductive structures, methods of forming capacitors, methods of forming an interconnect between a substrate bit line contact and a bit line in DRAM, and methods of forming DRAM memory cell
US20060271262A1 (en) * 2005-05-24 2006-11-30 Mclain Harry P Iii Wireless agricultural network
US20110003744A1 (en) * 2005-05-25 2011-01-06 Novo Nordisk A/S Glycopegylated Erythropoietin Formulations
JP5216580B2 (ja) * 2005-05-25 2013-06-19 ノヴォ ノルディスク アー／エス グリコペグ化第ｉｘ因子
WO2006133013A2 (en) * 2005-06-03 2006-12-14 The Regents Of The University Of California Methods of generating protein variants with altered function
KR101502920B1 (ko) 2005-06-21 2015-03-17 조마 (유에스) 엘엘씨 ＩＬ-１β 결합성 항체 및 그의 단편
KR20080025174A (ko) 2005-06-23 2008-03-19 메디뮨 인코포레이티드 응집 및 단편화 프로파일이 최적화된 항체 제제
US7582291B2 (en) * 2005-06-30 2009-09-01 The Rockefeller University Bacteriophage lysins for Enterococcus faecalis, Enterococcus faecium and other bacteria
EP1902136B1 (en) 2005-07-01 2012-01-18 Basf Se Herbicide-resistant sunflower plants, polynucleotides encoding herbicide-resistant acetohydroxyacid synthase large subunit proteins, and methods of use
EP1902135A1 (en) 2005-07-06 2008-03-26 CropDesign N.V. Plant yield improvement by ste20-like gene expression
MX2007016045A (es) 2005-07-08 2008-03-10 Novozymes As Variantes de subtilasa.
WO2007009082A1 (en) * 2005-07-12 2007-01-18 Codon Devices, Inc. Compositions and methods for biocatalytic engineering
WO2007008951A1 (en) * 2005-07-12 2007-01-18 Codon Devices, Inc. Compositions and methods for design of non-immunogenic proteins
WO2007011722A2 (en) 2005-07-15 2007-01-25 President And Fellows Of Harvard College Reaction discovery system
US20070015195A1 (en) 2005-07-18 2007-01-18 Pioneer Hi-Bred International, Inc. Modified FRT recombination site libraries and methods of use
ES2502415T3 (es) 2005-07-29 2014-10-03 Targeted Growth, Inc. Protección de proteínas KRP mutantes negativas dominantes contra la inhibición activa del complejo ciclina-CDK por parte de KRP de tipo salvaje
RU2413532C2 (ru) 2005-07-29 2011-03-10 Зольвай Фармасьютиклз Гмбх Способ получения стерилизованного порошкообразного панкреатина
AU2006280144B2 (en) * 2005-08-10 2012-06-14 Sanofi Pasteur Biologics, Llc Vaccination against dengue virus infection
SI1919503T1 (sl) 2005-08-10 2015-02-27 Macrogenics, Inc. Identifikacija in inĺ˝eniring protiteles z variantnimi fc regijami in postopki za njih uporabo
US11266607B2 (en) 2005-08-15 2022-03-08 AbbVie Pharmaceuticals GmbH Process for the manufacture and use of pancreatin micropellet cores
US9198871B2 (en) 2005-08-15 2015-12-01 Abbott Products Gmbh Delayed release pancreatin compositions
US20070105755A1 (en) 2005-10-26 2007-05-10 Neose Technologies, Inc. One pot desialylation and glycopegylation of therapeutic peptides
EP2360248A1 (en) 2005-08-24 2011-08-24 The Rockefeller University Ply-GBS mutant lysins
KR101362710B1 (ko) 2005-08-26 2014-02-13 노보자임스 아데니움 바이오테크 에이/에스 항균 활성을 지닌 폴리펩타이드 및 이들을 암호화한폴리뉴클레오타이드
US7541431B2 (en) * 2005-09-07 2009-06-02 Maine Medical Center Cristin/R-spondin ligands active in the Wnt signaling pathway and methods, compositions and kits relating thereto
CA2621267A1 (en) 2005-09-07 2007-03-15 Nugen Technologies, Inc. Improved nucleic acid amplification procedure
EP2573179A1 (en) 2005-09-15 2013-03-27 CropDesign N.V. Plants having increased yield and method for making the same
US8173613B2 (en) * 2005-09-22 2012-05-08 Maine Medical Center Modulation of mesenchymal and metastatic cell growth
EP2385110A3 (en) 2005-09-30 2011-11-16 Novozymes, Inc. Methods for enhancing the degradation or conversion of cellulosic material
US7312079B1 (en) 2005-10-06 2007-12-25 Lexicon Pharmaceuticals, Inc. Variants of FAM3C
US7470532B2 (en) 2005-10-19 2008-12-30 E.I. Du Pont De Nemours And Company Mortierella alpina C16/18 fatty acid elongase
DK2484202T3 (en) 2005-10-28 2017-09-11 Dow Agrosciences Llc NEW HERBICID RESISTANCE GENES
US20090048440A1 (en) * 2005-11-03 2009-02-19 Neose Technologies, Inc. Nucleotide Sugar Purification Using Membranes
EP2327770A1 (en) 2005-11-07 2011-06-01 CropDesign N.V. Use of Group I shaggy-like kinases for increasing salt-stress resistance and yield of plants
US8853492B2 (en) 2005-11-07 2014-10-07 Cropdesign N.V. Plants having improved growth characteristics and a method for making the same
AR056790A1 (es) 2005-11-08 2007-10-24 Cropdesign Nv Plantas con caracteristicas de crecimiento aumentado y un metodo para obtenerlas
US20070118920A1 (en) 2005-11-09 2007-05-24 Basf Agrochemical Products B.V. Herbicide-resistant sunflower plants, polynucleotides encoding herbicide-resistant acetohydroxyacid synthase large subunit proteins, and methods of use
US20070107084A1 (en) 2005-11-10 2007-05-10 Pioneer Hi-Bred International, Inc. Dof (DNA binding with one finger) sequences and methods of use
US20070199096A1 (en) 2005-11-14 2007-08-23 E.I. Du Pont De Nemours And Company Compositions and Methods for Altering Alpha- and Beta-Tocotrienol Content
AU2006318652A1 (en) * 2005-11-18 2007-05-31 Gloucester Pharmaceuticals, Inc. Metabolite derivatives of the HDAC inhibitor FK228
GB2432366B (en) * 2005-11-19 2007-11-21 Alligator Bioscience Ab A method for in vitro molecular evolution of protein function
GB0526664D0 (en) * 2005-11-30 2006-02-08 Plasticell Ltd Method
EP2157172A3 (en) 2005-12-01 2010-03-24 CropDesign N.V. Plants having improved growth characteristics and methods for making the same
AR057205A1 (es) 2005-12-01 2007-11-21 Athenix Corp Genes grg23 y grg51 que confieren resistencia a herbicidas
EP2341140B1 (en) 2005-12-01 2017-07-19 Nuevolution A/S Enzymatic encoding methods for efficient synthesis of large libraries
AU2006320275B2 (en) * 2005-12-02 2012-06-07 Synthetic Genomics, Inc. Synthesis of error-minimized nucleic acid molecules
KR100784478B1 (ko) * 2005-12-05 2007-12-11 한국과학기술원 기능요소의 동시 삽입에 의한 신기능을 갖는 단백질을제조하는 방법
US20080313769A9 (en) 2006-01-12 2008-12-18 Athenix Corporation EPSP synthase domains conferring glyphosate resistance
EP2216403A3 (en) 2006-02-02 2010-11-24 Verenium Corporation Esterases and related nucleic acids and methods
NZ595498A (en) 2006-02-10 2013-05-31 Verenium Corp Cellulolytic enzymes, nucleic acids encoding them and methods for making and using them
ES2682284T3 (es) 2006-02-14 2018-09-19 Bp Corporation North America Inc. Xilanasas, ácidos nucleicos que las codifican y métodos para elaborarlas y usarlas
WO2007103768A2 (en) 2006-03-02 2007-09-13 Athenix Corporation Methods and compositions for improved enzyme activity in transgenic plant
EP2650306A1 (en) 2006-03-06 2013-10-16 Aeres Biomedical Limited Humanized Anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP2316962B1 (en) * 2006-03-07 2014-07-09 Cargill, Incorporated Aldolases, nucleic acids encoding them and methods for making and using them
CA2645225A1 (en) 2006-03-07 2007-09-13 Verenium Corporation Aldolases, nucleic acids encoding them and methods for making and using them
US20070214515A1 (en) 2006-03-09 2007-09-13 E.I.Du Pont De Nemours And Company Polynucleotide encoding a maize herbicide resistance gene and methods for use
ATE513907T1 (de) 2006-03-20 2011-07-15 Novozymes Inc Polypeptide mit endoglucanase-aktivität und polynukleotide zu ihrer kodierung
US8703278B2 (en) * 2006-03-28 2014-04-22 Honeywell International Inc. Light weight printed wiring board
EP2374877A3 (en) 2006-03-30 2012-11-07 Novozymes A/S Polypeptides having endoglucanase activity and polynucleotides encoding same
MX2008010669A (es) 2006-03-31 2008-09-03 Basf Plant Science Gmbh Plantas con caracteristicas mejoradas y un metodo para prepararlas.
EP1842915A1 (en) 2006-04-04 2007-10-10 Libragen Method of in vitro polynucleotide sequences shuffling by recursive circular DNA molecules fragmentation and ligation
PL2365064T3 (pl) 2006-04-04 2015-05-29 Novozymes As Warianty fitazy
BRPI0710455A2 (pt) 2006-04-19 2011-08-16 Pioneer Hi Bred Int polipeptìdeo isolado, polinucleotìdeo isolado e métodos para aumentar o nìvel de um polipeptìdeo em uma planta, para modular o nìvel de um polipeptìdeo em uma planta e para modificar o crescimento de uma planta
WO2008013589A2 (en) * 2006-04-24 2008-01-31 Gloucester Pharmaceuticals Treatment of ras-expressing tumors
CN101490265B (zh) 2006-05-16 2013-09-04 先锋高级育种国际公司 抗真菌多肽
EP2018435B1 (en) 2006-05-17 2012-07-11 Pioneer Hi-Bred International Inc. Artificial plant minichromosomes
US20090087840A1 (en) * 2006-05-19 2009-04-02 Codon Devices, Inc. Combined extension and ligation for nucleic acid assembly
US10072256B2 (en) 2006-05-22 2018-09-11 Abbott Products Gmbh Process for separating and determining the viral load in a pancreatin sample
US20090241218A1 (en) 2006-05-30 2009-09-24 Cropdesign N.V. Plants having enhanced yield-related traits and a method for making the same
US8273952B2 (en) 2006-06-08 2012-09-25 Basf Plant Science Gmbh Plants having improved growth characteristics and method for making the same
US8957027B2 (en) * 2006-06-08 2015-02-17 Celgene Corporation Deacetylase inhibitor therapy
CN105087634A (zh) 2006-06-15 2015-11-25 克罗普迪塞恩股份有限公司 具有增强的产量相关性状的nac转录因子受调节表达的植物和用于产生该植物的方法
WO2007144190A2 (en) 2006-06-15 2007-12-21 Cropdesign N.V. Plants with modulated expression of nac transcription factors having enhanced yield-related traits and a method for making the same
EP2455393A3 (en) 2006-06-15 2012-08-22 Athenix Corporation A family of pesticidal proteins and methods for their use
CA2997344C (en) 2006-06-19 2023-04-04 22Nd Century Limited, Llc Nucleic acid encoding n-methylputrescine oxidase and uses thereof
JP2009541333A (ja) * 2006-06-23 2009-11-26 クインテセンス バイオサイエンシーズ インコーポレーティッド 修飾リボヌクレアーゼ
WO2008019199A2 (en) 2006-06-26 2008-02-14 Macrogenics, Inc. FCγRIIB-SPECIFIC ANTIBODIES AND METHODS OF USE THEREOF
US20080004410A1 (en) * 2006-06-30 2008-01-03 Yu-Chin Lai Hydrophilic macromonomers having alpha,beta-conjugated carboxylic terminal group and medical devices incorporating same
US7572618B2 (en) 2006-06-30 2009-08-11 Bristol-Myers Squibb Company Polynucleotides encoding novel PCSK9 variants
US7977535B2 (en) 2006-07-12 2011-07-12 Board Of Trustees Of Michigan State University DNA encoding ring zinc-finger protein and the use of the DNA in vectors and bacteria and in plants
ES2584506T3 (es) 2006-07-13 2016-09-28 Dsm Ip Assets B.V. Uso de amilasas bacterianas en pienso para animales bovinos
EP2049151A4 (en) 2006-07-17 2010-03-24 Quintessence Biosciences Inc METHOD AND COMPOSITIONS FOR THE TREATMENT OF CANCER
JP2009544327A (ja) * 2006-07-21 2009-12-17 ノヴォ ノルディスク アー／エス Ｏ−結合型グリコシル化配列によるペプチドのグリコシル化
EP2054511A1 (en) * 2006-07-25 2009-05-06 Commonwealth Scientific and Industrial Research Organization Identification of a novel type of sucrose synthase and use thereof in fiber modification
EP2189533A1 (en) 2006-08-02 2010-05-26 CropDesign N.V. Plants having improved characteristics and a method for making the same
EP2546262A1 (en) 2006-08-02 2013-01-16 CropDesign N.V. Modifying the content of storage compounds in seeds by expression of a Class II HD-Zip transcription factor
EP2444413A1 (en) 2006-08-04 2012-04-25 Verenium Corporation Methods for oil or gas well drilling, washing and/or fracturing
US8252559B2 (en) * 2006-08-04 2012-08-28 The California Institute Of Technology Methods and systems for selective fluorination of organic molecules
US8026085B2 (en) * 2006-08-04 2011-09-27 California Institute Of Technology Methods and systems for selective fluorination of organic molecules
US7968691B2 (en) * 2006-08-23 2011-06-28 Danisco Us Inc. Pullulanase variants with increased productivity
KR101314362B1 (ko) 2006-08-28 2013-10-10 라 졸라 인스티튜트 포 앨러지 앤드 이뮤놀로지 길항성 인간 ｌｉｇｈｔ-특이적 인간 모노클로날 항체
PE20081216A1 (es) 2006-09-01 2008-09-04 Therapeutic Human Polyclonals Inc Expresion mejorada de la inmunoglobulina humana o humanizada en animales transgenicos no humanos
EP2061904A2 (en) 2006-09-06 2009-05-27 Ortho-McNeil Pharmaceutical, Inc. Biomarkers for assessing response to c-met treatment
AU2007294575B2 (en) 2006-09-08 2013-06-27 Viela Bio, Inc. Humanized anti-CD19 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP2617819B1 (en) 2006-09-21 2016-04-27 BASF Enzymes LLC Phytases, nucleic acids encoding them and methods for making and using them
BRPI0716872A2 (pt) 2006-09-21 2015-06-16 Verenium Corp Fosfolipases, ácidos nucléicos codificando-as e métodos para fabricação e uso delas
CN104938794A (zh) 2006-09-29 2015-09-30 诺维信公司 用于动物饲料的木聚糖酶
JP2010505874A (ja) 2006-10-03 2010-02-25 ノヴォ ノルディスク アー／エス ポリペプチドコンジュゲートの精製方法
LT2068907T (lt) * 2006-10-04 2018-01-10 Novo Nordisk A/S Glicerolio sujungti pegilinti sacharidai ir glikopeptidai
GB0620715D0 (en) 2006-10-18 2006-11-29 Univ Durham Ethanol production
US20100162433A1 (en) 2006-10-27 2010-06-24 Mclaren James Plants with improved nitrogen utilization and stress tolerance
EP2433635A1 (en) 2006-11-10 2012-03-28 Massachusetts Institute Of Technology PAK Modulators
WO2008133722A2 (en) * 2006-11-10 2008-11-06 Ucb Pharma S.A. Anti human sclerostin antibodies
EP2097450A2 (en) * 2006-11-10 2009-09-09 Amgen Inc. Antibody-based diagnostics and therapeutics
WO2008059048A1 (en) 2006-11-16 2008-05-22 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same using consensus sequences from the yabby protein family
MX2009005280A (es) 2006-11-24 2009-08-12 Cropdesign Nv Plantas transgenicas que comprenden como transgen un tcp clase i o clavata 1 (clv1) o polipeptido de cah3 que tienen un rendimiento aumentado de semillas y un metodo para generar las mismas.
DK2089516T3 (da) 2006-11-29 2012-02-20 Novozymes Inc Fremgangsmåder til at forbedre indføringen af DNA i bakterielle celler
EP2099818A2 (en) 2006-11-29 2009-09-16 Novozymes Inc. Bacillus licheniformis chromosome
EP2610334B1 (en) * 2006-12-07 2016-10-26 Dow AgroSciences LLC Novel selectable marker genes
WO2008073914A2 (en) 2006-12-10 2008-06-19 Dyadic International Inc. Expression and high-throughput screening of complex expressed dna libraries in filamentous fungi
US9862956B2 (en) 2006-12-10 2018-01-09 Danisco Us Inc. Expression and high-throughput screening of complex expressed DNA libraries in filamentous fungi
EP2487244A1 (en) 2006-12-15 2012-08-15 CropDesign N.V. Plants having enhanced yield-related traits and a method for making the same
WO2008074116A1 (en) 2006-12-20 2008-06-26 Alellyx S.A. Nucleic acid molecules encoding plant proteins in the c3hc4 family and methods for the alteration of plant cellulose and lignin content
EP2094852B1 (en) 2006-12-20 2014-01-22 Monsanto do Brasil LTDA Nucleic acid constructs and methods for altering plant fiber length and/or plant height
CN102676575B (zh) 2006-12-21 2015-09-30 巴斯福植物科学有限公司 具有增强的产量相关性状的植物和用于产生该植物的方法
PL3101128T3 (pl) 2006-12-21 2019-09-30 Basf Enzymes Llc Amylazy i glukoamylazy, kwasy nukleinowe kodujące te związki oraz sposoby wytwarzania tych związków oraz stosowania ich
CN107090482A (zh) * 2006-12-29 2017-08-25 细胞基因公司 制备Romidepsin
MX2009006966A (es) * 2006-12-29 2009-12-09 Gloucester Pharmaceuticals Inc Tratamientos para cancer a base de romidepsina.
DE102007003143A1 (de) 2007-01-16 2008-07-17 Henkel Kgaa Neue Alkalische Protease aus Bacillus gibsonii und Wasch- und Reinigungsmittel enthaltend diese neue Alkalische Protease
NZ610301A (en) 2007-01-30 2015-03-27 Bp Corp North America Inc Enzymes for the treatment of lignocellulosics, nucleic acids encoding them and methods for making and using them
AR065100A1 (es) 2007-01-30 2009-05-13 Cropdesign Nv Plantas con rasgos relacionados con el rendimiento mejorado y un metodo para producirlas
EP2275560A3 (en) 2007-01-31 2012-01-25 BASF Plant Science GmbH Trangenic plants wherein the transgene is a memeber of the REMORIN gene family or a consensus sequence thereof having enhanced yield-related traits and a method for making the same
US20080248545A1 (en) * 2007-02-02 2008-10-09 The California Institute Of Technology Methods for Generating Novel Stabilized Proteins
US8143046B2 (en) 2007-02-07 2012-03-27 Danisco Us Inc., Genencor Division Variant Buttiauxella sp. phytases having altered properties
US20080268517A1 (en) * 2007-02-08 2008-10-30 The California Institute Of Technology Stable, functional chimeric cytochrome p450 holoenzymes
ATE518946T1 (de) 2007-02-08 2011-08-15 Codexis Inc Ketoreduktasen und anwendungen davon
EP2118281A2 (en) 2007-02-12 2009-11-18 Codexis, Inc. Structure-activity relationships
CN101631868B (zh) 2007-02-16 2016-02-10 巴斯福植物科学有限公司 用于在单子叶植物中调节胚特异性表达的核酸序列
AR065552A1 (es) 2007-02-28 2009-06-17 Crop Functional Genomics Ct Plantas con rasgos aumentados relacionados con el rendimiento y un metodo para producirlas
WO2008118445A1 (en) * 2007-03-26 2008-10-02 Promega Corporation Methods to quench light from optical reactions
DK2129781T3 (da) 2007-03-26 2014-03-31 Novozymes As Hafnia-phytase
KR101540823B1 (ko) 2007-03-30 2015-07-30 메디뮨 엘엘씨 항체 제제
DE102007016139A1 (de) 2007-03-30 2008-10-02 Jenabios Gmbh Verfahren zur regioselektiven Oxygenierung von N-Heterozyklen
KR20150064246A (ko) * 2007-04-03 2015-06-10 바이오제너릭스 게엠베하 글리코페길화 ｇ―ｃｓｆ를 이용하는 치료 방법
KR20100016165A (ko) 2007-04-04 2010-02-12 바스프 에스이 제초제-저항성 브라시카 식물 및 사용 방법
EP2132292B1 (en) 2007-04-09 2014-10-08 Novozymes A/S An enzymatic treatment of skin and hide degreasing
CA2900183A1 (en) 2007-05-03 2008-11-13 Yves Hatzfeld Plants having enhanced yield-related traits and a method for making the same
EP2164868B1 (en) 2007-05-04 2015-03-25 Technophage, Investigação E Desenvolvimento Em Biotecnologia, SA Engineered rabbit antibody variable domains and uses thereof
RU2549701C2 (ru) 2007-05-07 2015-04-27 Медиммун, Ллк Анти-icos антитела и их применение в лечении онкологических, связанных с трансплантацией и аутоиммунных заболеваний
US8298781B2 (en) * 2007-05-08 2012-10-30 Duke University Compositions and methods for characterizing and regulating olfactory sensation
EP2145007B1 (en) 2007-05-09 2017-01-25 Dow AgroSciences LLC Novel herbicide resistance genes
WO2008141190A1 (en) * 2007-05-11 2008-11-20 Wisconsin Alumni Research Foundation Heterologous production of capreomycin and generation of new capreomycin derivatives through metabolic engineering
PL2068927T3 (pl) 2007-05-14 2016-06-30 Medimmune Llc Sposoby redukcji poziomów eozynofilii
BRPI0811185A2 (pt) 2007-05-23 2014-10-07 Cropdesign Nv Método para intensificação de características relacionadas a rendimento em plantas com relação às plantas de controle, planta, parte de planta ou célula de planta, construto, uso de um construto, método para a produção de uma planta transgênica tendo características relacionadas a rendimento intensificadas com relação às plantas de controle, planta transgênica, partes colhíveis de uma planta, produtos derivados de uma planta, e, uso de uma seqüência de ácido nucleico
CN110885834B (zh) 2007-05-25 2023-08-11 22世纪有限责任公司 编码调节生物碱合成之转录因子的核酸序列及其在改良植物代谢中的应用
EP2152733A2 (en) 2007-05-25 2010-02-17 CropDesign N.V. Yield enhancement in plants by modulation of maize alfins
EP2162535A4 (en) * 2007-06-04 2011-02-23 Novo Nordisk As O-linked glycosylation using N-acetylglucosamine transferases
JP5876649B2 (ja) 2007-06-12 2016-03-02 ラツィオファルム ゲーエムベーハーｒａｔｉｏｐｈａｒｍ ＧｍｂＨ ヌクレオチド糖の改良製造法
US7625555B2 (en) 2007-06-18 2009-12-01 Novagen Holding Corporation Recombinant human interferon-like proteins
EP2158221B1 (en) 2007-06-21 2018-08-29 MacroGenics, Inc. Covalent diabodies and uses thereof
WO2009003977A2 (en) 2007-06-29 2009-01-08 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same
US7968811B2 (en) * 2007-06-29 2011-06-28 Harley-Davidson Motor Company Group, Inc. Integrated ignition and key switch
WO2009012357A2 (en) 2007-07-17 2009-01-22 The General Hospital Corporation Methods to identify and enrich for populations of ovarian cancer stem cells and somatic ovarian stem cells and uses thereof
EP2173884B1 (en) 2007-07-20 2013-09-18 BASF Plant Science GmbH Plants having increased yield-related traits and a method for making the same
AR067748A1 (es) 2007-07-31 2009-10-21 Basf Plant Science Gmbh Plantas que tienen rasgos mejorados relacionados con el rendimiento y un metodo para obtenerlas
CN101952441B (zh) 2007-07-31 2013-08-14 巴斯夫植物科学有限公司 具有增强的产量相关性状的植物及其制备方法
US7923236B2 (en) 2007-08-02 2011-04-12 Dyadic International (Usa), Inc. Fungal enzymes
CN101842159B (zh) * 2007-08-09 2014-09-24 赛路拉公司 关联的多参数单细胞测定及残余生物材料回收的方法和装置
WO2009029554A2 (en) 2007-08-24 2009-03-05 Codexis, Inc. Improved ketoreductase polypeptides for the stereoselective production of (r)-3-hydroxythiolane
US8207112B2 (en) * 2007-08-29 2012-06-26 Biogenerix Ag Liquid formulation of G-CSF conjugate
WO2009033071A2 (en) 2007-09-07 2009-03-12 Dyadic International, Inc. Novel fungal enzymes
US7749708B2 (en) * 2007-09-12 2010-07-06 Transgenomic, Inc. Method for identifying the sequence of one or more variant nucleotides in a nucleic acid molecule
US7579155B2 (en) * 2007-09-12 2009-08-25 Transgenomic, Inc. Method for identifying the sequence of one or more variant nucleotides in a nucleic acid molecule
WO2009036379A2 (en) 2007-09-14 2009-03-19 Adimab, Inc. Rationally designed, synthetic antibody libraries and uses therefor
US8877688B2 (en) 2007-09-14 2014-11-04 Adimab, Llc Rationally designed, synthetic antibody libraries and uses therefor
WO2009034188A1 (en) 2007-09-14 2009-03-19 Basf Plant Science Gmbh Plants having increased yield-related traits and a method for making the same
CL2008002775A1 (es) * 2007-09-17 2008-11-07 Amgen Inc Uso de un agente de unión a esclerostina para inhibir la resorción ósea.
WO2009037253A2 (en) 2007-09-18 2009-03-26 Novozymes A/S Polypeptides having tyrosinase activity and polynucleotides encoding same
JP2010538670A (ja) 2007-09-21 2010-12-16 ビーエーエスエフ プラント サイエンス ゲーエムベーハー 増強された収穫高関連形質を有する植物およびその作製方法
CN110577945A (zh) 2007-10-03 2019-12-17 维莱尼姆公司 木聚糖酶、编码它们的核酸以及其制备和应用方法
JP2010540681A (ja) * 2007-10-08 2010-12-24 クインテッセンス バイオサイエンシズ，インコーポレーテッド リボヌクレアーゼに基づく治療のための組成物及び方法
CN101821395B (zh) 2007-10-10 2013-11-06 阿森尼克斯公司 编码cry1ac的合成基因
WO2009052242A2 (en) 2007-10-16 2009-04-23 Athenix Corporation Axmi-066 and axmi-076: delta-endotoxin proteins and methods for their use
DE102007051092A1 (de) 2007-10-24 2009-04-30 Henkel Ag & Co. Kgaa Subtilisin aus Becillus pumilus und Wasch- und Reinigungsmittel enthaltend dieses neue Subtilisin
WO2009056566A2 (en) 2007-10-29 2009-05-07 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same
US7618801B2 (en) * 2007-10-30 2009-11-17 Danison US Inc. Streptomyces protease
US7871802B2 (en) 2007-10-31 2011-01-18 E.I. Du Pont De Nemours And Company Process for enzymatically converting glycolonitrile to glycolic acid
US7741088B2 (en) * 2007-10-31 2010-06-22 E.I. Dupont De Nemours And Company Immobilized microbial nitrilase for production of glycolic acid
DK2215202T3 (da) 2007-11-05 2017-11-27 Danisco Us Inc VARIANTER AF BACILLUS sp. TS-23 ALPHA-AMYLASE MED ÆNDREDE EGENSKABER
BRPI0819184B1 (pt) 2007-11-05 2022-05-10 Danisco Us Inc Variantes de alfa-amilase com propriedades alteradas, polinucleotídeo, vetor, célula hospedeira, composição, kit, bem como métodos para tratamento de um caldo de amido, para produção de um substrato fermentável, e para tratamento de um material contendo amido
CA2706506A1 (en) 2007-11-22 2009-05-28 Valerie Frankard Plants having increased yield-related traits and a method for making the same
EP2574621A1 (en) 2007-11-26 2013-04-03 BASF Plant Science GmbH Plants having enhanced yield-related traits and a method for making the same
BRPI0820552A2 (pt) 2007-11-27 2014-10-14 Novozymes As Polipetídeo isolado, polinucleotídeo isolado, construção de ácido nucléico, vetor de expressão recombinante, célula hospedeira recombinante, métodos para produzir o polipeptídeo, para produzir uma proteína, e para degradar ou converter um material de biomassa, planta, parte vegetal ou célula vegetal transgêncica, método, composição, e. uso do polipeptídio.
CA2706639A1 (en) 2007-11-29 2009-06-04 Novozymes A/S Synthase inhibitor screening method
CN104650235A (zh) 2007-11-30 2015-05-27 葛兰素集团有限公司 抗原结合构建体
CN101909461B (zh) 2007-12-06 2015-10-07 诺维信公司 具有乙酰木聚糖酯酶活性的多肽和编码该多肽的多核苷酸
US8318481B2 (en) * 2007-12-07 2012-11-27 Pfenex Inc. High copy number self-replicating plasmids in pseudomonas
KR20100101656A (ko) * 2007-12-14 2010-09-17 암젠 인크 항-스클레로스틴 항체를 사용한 골절 치료 방법
WO2009085935A2 (en) 2007-12-19 2009-07-09 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
CN101969759A (zh) 2007-12-20 2011-02-09 巴斯夫植物科学有限公司 具有增强的产量相关性状的植物及其制备方法
BRPI0821253A2 (pt) 2007-12-21 2015-06-16 Danisco Método de degomagem, método de tratamento de uma fase de goma e uso de lipídio aciltransferase na degomagem com água de um óleo comestível
EP2238261B1 (en) 2008-01-03 2013-12-04 Verenium Corporation Isomerases, nucleic acids encoding them and methods for making and using them
CN104651381A (zh) 2008-01-03 2015-05-27 巴斯夫酶有限责任公司 转移酶和氧化还原酶、编码它们的核酸以及其制备和应用方法
WO2009089396A2 (en) * 2008-01-08 2009-07-16 Neose Technologies, Inc. Glycoconjugation of polypeptides using oligosaccharyltransferases
ES2613963T3 (es) 2008-01-18 2017-05-29 Medimmune, Llc Anticuerpos manipulados con cisteína para conjugación específica de sitio
BRPI0906617A2 (pt) 2008-01-25 2015-07-14 Basf Plant Science Gmbh Método para melhorar características relacionadas ao rendimento em plantas com relação às plantas controle, construto, uso de um construto, planta, parte da planta ou célula de planta, método para a produção de uma planta transgênica, planta transgênica, partes colhíveis de uma planta, produtos, e, uso de um ácido nucléico.
CN101978064A (zh) 2008-01-31 2011-02-16 北京生命科学研究所 具有改变的生长和/或发育的植物及其生产方法
RU2526516C2 (ru) 2008-02-04 2014-08-20 ДАНИСКО ЮЭс ИНК. Варианты альфа-амилазы ts-23 с измененными свойствами
US20100049602A1 (en) * 2008-02-07 2010-02-25 Softky William R Systems and Methods for Measuring the Effectiveness of Advertising
PT2250279E (pt) 2008-02-08 2016-06-03 Medimmune Llc Anticorpos anti-ifnár1 com afinidade reduzida para o ligando fc
EP2088432A1 (en) * 2008-02-11 2009-08-12 MorphoSys AG Methods for identification of an antibody or a target
WO2009102899A1 (en) 2008-02-12 2009-08-20 Codexis, Inc. Method of generating an optimized diverse population of variants
EP2250594B1 (en) * 2008-02-12 2017-04-19 Codexis, Inc. Method of generating an optimized, diverse population of variants
US8034568B2 (en) 2008-02-12 2011-10-11 Nugen Technologies, Inc. Isothermal nucleic acid amplification methods and compositions
CN103497246B (zh) 2008-02-27 2016-08-10 诺沃—诺迪斯克有限公司 缀合的因子viii分子
GB2470672B (en) 2008-03-21 2012-09-12 Nugen Technologies Inc Methods of RNA amplification in the presence of DNA
US20110020368A1 (en) 2008-03-25 2011-01-27 Nancy Hynes Treating cancer by down-regulating frizzled-4 and/or frizzled-1
US20110020830A1 (en) * 2008-03-31 2011-01-27 Schneider Jane C Design for rapidly cloning one or more polypeptide chains into an expression system
BRPI0910576A2 (pt) 2008-04-16 2015-08-04 Basf Plant Science Gmbh Métodos para intensificar e/ou aumentar características relacionadas com rendimento em plantas, e para a produção de uma planta transgênica apresentando rendimento aumentado, construção, usos de uma construção e de um ácido nucleico que codifica um polipeptídeo de ornitina descarboxilase, planta, parte de planta ou célula de planta, planta transgênica, partes colhíveis de uma planta, e, produtos.
DK2281034T3 (en) 2008-04-30 2016-01-25 Dupont Nutrition Biosci Aps A method using alcohol dehydrogenase Pseudoglucanobacter saccharoketogenes
PT2274331E (pt) 2008-05-02 2014-02-27 Novartis Ag Moléculas de ligação baseadas em fibronectina melhoradas e suas utilizações
JP2011521626A (ja) 2008-05-02 2011-07-28 エピセンター テクノロジーズ コーポレーション Ｒｎａポリホスファターゼの組成物、キットおよびそれらの使用
CA2722806A1 (en) 2008-05-05 2009-11-12 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same
WO2009136293A2 (en) 2008-05-08 2009-11-12 Alellyx Applied Genomics Genes and methods for increasing disease resistance in plants
JP2010043063A (ja) 2008-05-09 2010-02-25 Agency For Science Technology & Research 川崎病の診断及び治療
WO2009143397A2 (en) 2008-05-23 2009-11-26 E. I. Du Pont De Nemours And Company Novel dgat genes for increased seed storage lipid production and altered fatty acid profiles in oilseed plants
WO2009147201A2 (en) * 2008-06-04 2009-12-10 Modiquest B.V. Anti-inflammatory agents
BRPI0913570A2 (pt) 2008-06-06 2015-12-15 Procter & Gamble composição detergente compreendendo uma variante de uma xiloglucanase da família 44
EP2674488A1 (en) 2008-06-06 2013-12-18 Novozymes A/S Variants of a family 44 xyloglucanase
AU2009256280B2 (en) 2008-06-06 2013-03-07 Danisco Us Inc. Geobacillus stearothermophilus alpha-amylase (AmyS) variants with improved properties
WO2009152336A1 (en) 2008-06-13 2009-12-17 Codexis, Inc. Method of synthesizing polynucleotide variants
US20090312196A1 (en) 2008-06-13 2009-12-17 Codexis, Inc. Method of synthesizing polynucleotide variants
US8383346B2 (en) 2008-06-13 2013-02-26 Codexis, Inc. Combined automated parallel synthesis of polynucleotide variants
MX2010012720A (es) 2008-06-20 2010-12-14 Basf Plant Science Gmbh Plantas que tienen rasgos mejorados relacionados con el rendimiento y un metodo para obtenerlas.
MX2010014500A (es) 2008-06-24 2011-05-30 Codexis Inc Procedimientos biocataliticos para la preparacion de compuestos de prolina biciclica fusionada sustancialmente estereomericamente puros.
MX2010013910A (es) 2008-06-25 2011-06-20 Athenix Corp Genes de toxinas y metodos para su uso.
WO2009156360A1 (en) 2008-06-26 2009-12-30 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same
EP2297190A2 (en) 2008-07-02 2011-03-23 Athenix Corporation Axmi-115, axmi-113, axmi-005, axmi-163 and axmi-184: vip3a insecticidal proteins from bacillus thuringiensis and methods for their use
US8748699B2 (en) 2008-07-04 2014-06-10 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same by overexpressing a polynucleotide encoding a TFL1-like protein
US9175303B2 (en) 2008-07-17 2015-11-03 Basf Plant Science Gmbh Plants having enhanced yield-related traits and a method for making the same
EA201100910A1 (ru) 2008-07-18 2012-05-30 Новозимс Эдениум Байотек А/С Лечение воспалительных заболеваний кишечника бета-дефенсинами млекопитающих
WO2010009391A1 (en) 2008-07-18 2010-01-21 Bristol-Myers Squibb Company Compositions monovalent for cd28 binding and methods of use
US20100034897A1 (en) * 2008-07-22 2010-02-11 Northwestern University Method for Removal or Inactivation of Heparin
AU2009275889A1 (en) 2008-07-31 2010-02-04 Basf Plant Science Gmbh Plants having modified growth characteristics and a method for making the same
NZ590634A (en) 2008-08-05 2012-07-27 Novartis Ag Compositions and methods for antibodies targeting complement protein c5
EP2315774A1 (en) 2008-08-20 2011-05-04 BASF Plant Science GmbH Plants having enhanced yield-related traits and a method for making the same
HUE026181T2 (en) 2008-08-27 2016-05-30 Codexis Inc Ketoreductase polypeptide for the preparation of 3-aryl-3-hydroxypropanamine from 3-aryl-3-ketopropanamine
WO2010025287A2 (en) 2008-08-27 2010-03-04 Codexis, Inc. Ketoreductase polypeptides for the production of 3-aryl-3-hydroxypropanamine from a 3-aryl-3-ketopropanamine
WO2010027710A2 (en) 2008-08-27 2010-03-11 Codexis, Inc. Ketoreductase polypeptides and uses thereof
US8357503B2 (en) 2008-08-29 2013-01-22 Bunge Oils, Inc. Hydrolases, nucleic acids encoding them and methods for making and using them
US8198062B2 (en) 2008-08-29 2012-06-12 Dsm Ip Assets B.V. Hydrolases, nucleic acids encoding them and methods for making and using them
US8153391B2 (en) 2008-08-29 2012-04-10 Bunge Oils, Inc. Hydrolases, nucleic acids encoding them and methods for making and using them
CN102186972B (zh) 2008-08-29 2014-08-20 科德克希思公司 用于立体选择性生产(4s)-3-[(5s)-5-(4-氟苯基)-5-羟基戊酰基]-4-苯基-1，3-噁唑烷-2-酮的酮还原酶多肽
WO2010033229A2 (en) * 2008-09-22 2010-03-25 Calmune Corporation Methods and vectors for display of molecules and displayed molecules and collections
WO2010033237A2 (en) * 2008-09-22 2010-03-25 Calmune Corporation Methods for creating diversity in libraries and libraries, display vectors and methods, and displayed molecules
EP2716762A3 (en) 2008-09-24 2014-08-13 BASF Plant Science GmbH Plants having enhanced yield-related traits and a method for making the same
WO2010036515A1 (en) 2008-09-25 2010-04-01 Danisco Us Inc. Alpha-amylase blends and methods for using said blends
EP2733212B1 (en) 2008-09-26 2018-09-05 BASF Agrochemical Products, B.V. Herbicide-resistant ahas-mutants and methods of use
MX2011003132A (es) 2008-09-26 2011-04-21 Novozymes As Variantes de fitasa de hafnia.
WO2010045002A2 (en) 2008-09-26 2010-04-22 Tocagen Inc. Gene therapy vectors and cytosine deaminases
JP2012504423A (ja) 2008-10-01 2012-02-23 クインテッセンス バイオサイエンシズ，インコーポレーテッド 治療用リボヌクレアーゼ
US8337905B2 (en) 2008-10-03 2012-12-25 E. I. Du Pont De Nemours And Company Multi-component peracid generation system
WO2010052244A1 (en) * 2008-11-05 2010-05-14 Morphosys Ag Deconvolution method
AU2009313560B2 (en) 2008-11-07 2016-04-14 Fabrus Llc Combinatorial antibody libraries and uses thereof
ES2719496T3 (es) 2008-11-12 2019-07-10 Medimmune Llc Formulación de anticuerpo
BRPI0921061A2 (pt) 2008-11-12 2016-07-19 Basf Plant Science Gmbh métodos para intensificar tolerância a estresse abiótico e/ou intensificar características relacionadas ao rendimento em plantas com relação a uma planta de controle, e para produzir uma planta transgênica com tolerância a estresse abiótico e/ou características relacionadas ao rendimento para uma planta de controle, partes colhíveis, produtos, construto, planta, parte de planta, ou célula de planta, usos de um construto, e de um ácido nucleico, molécula de ácido nucleico isolada, e, polipeptídeo isolado
WO2010059413A2 (en) 2008-11-20 2010-05-27 Novozymes, Inc. Polypeptides having amylolytic enhancing activity and polynucleotides encoding same
CA2745074A1 (en) 2008-12-03 2010-06-10 Basf Plant Science Gmbh Arp6 polypeptide-expressing plants having enhanced abiotic stress tolerance and/or enhanced yield-related traits and a method for making the same
CN102300986A (zh) 2008-12-04 2011-12-28 诺维信股份有限公司 具有纤维素分解增强活性的多肽和编码该多肽的多核苷酸
WO2010065867A1 (en) 2008-12-04 2010-06-10 Pioneer Hi-Bred International, Inc. Methods and compositions for enhanced yield by targeted expression of knotted1
US9493759B2 (en) 2008-12-12 2016-11-15 Novozymes, Inc. Polypeptides having aspartic endopeptidase activity and polynucleotides encoding same
WO2010068650A1 (en) 2008-12-12 2010-06-17 Novozymes, Inc. Polypeptides having lipase activity and polynucleotides encoding same
CN102369280B (zh) 2008-12-15 2015-02-11 阿森尼克斯公司 编码线虫毒素的基因
WO2010074972A1 (en) 2008-12-15 2010-07-01 Novozymes, Inc. Polypeptides having catalase activity and polynucleotides encoding same
US8633030B2 (en) 2008-12-16 2014-01-21 Novozymes, Inc. Polypeptides having carboxypeptidase activity and polynucleotides encoding same
US20110271407A1 (en) 2008-12-16 2011-11-03 Novozymes, Inc. Polypeptides Having Alpha-Mannosidase Activity And Polynucleotides Encoding Same
DE112009003749T5 (de) 2008-12-17 2012-11-15 Basf Plant Science Gmbh Pflanzen mit gesteigerten ertragsbezogenen Eigenschaften und/oder gesteigerter abiotischerStresstoleranz und Verfahren zur Herstellung derselben
WO2010070110A2 (en) 2008-12-19 2010-06-24 Novozymes A/S Use of enzymes having silicase activity
KR101940059B1 (ko) 2008-12-19 2019-01-18 마크로제닉스, 인크. 공유결합형 디아바디 및 이의 용도
EP2671951A3 (en) 2008-12-22 2014-06-04 Athenix Corporation Pesticidal genes from Brevibacillus and methods for their use
ES2534581T3 (es) 2008-12-23 2015-04-24 Athenix Corporation Gen AXMI-150 de la delta-endotoxina y métodos para su uso
US8329438B2 (en) * 2008-12-25 2012-12-11 Codexis, Inc. Enone reductases
HUE052297T2 (hu) 2009-01-08 2021-04-28 Codexis Inc Transzamináz polipeptidek
EP2206723A1 (en) 2009-01-12 2010-07-14 Bonas, Ulla Modular DNA-binding domains
US20110239315A1 (en) 2009-01-12 2011-09-29 Ulla Bonas Modular dna-binding domains and methods of use
EP2379720B1 (en) 2009-01-20 2016-08-17 Alona Zilberberg Mir-21 promoter driven targeted cancer therapy
AU2010206181B2 (en) 2009-01-21 2015-01-15 Novozymes A/S Polypeptides having esterase activity and nucleic acids encoding the same
BRPI1007465A2 (pt) 2009-01-28 2015-11-24 Basf Plant Science Co Gmbh método para realçar traços relacionados com o rendimento em plantas em relação às plantas de controle, para produzir uma planta transgênica tendo rendimento aumentado, molécula de ácido nucleico isolada, polipeptídio isolado, construçao, uso de uma construção e de um ácido nucleico, célula de planta, e, produtos
WO2010088387A1 (en) 2009-01-28 2010-08-05 Novozymes, Inc. Polypeptides having beta-glucosidase activity and polynucleotides encoding same
WO2010088463A2 (en) 2009-01-30 2010-08-05 Novozymes, Inc. Polypeptides having expansin activity and polynucleotides encoding same
WO2010088447A1 (en) 2009-01-30 2010-08-05 Novozymes A/S Polypeptides having alpha-amylase activity and polynucleotides encoding same
US8852608B2 (en) 2009-02-02 2014-10-07 Medimmune, Llc Antibodies against and methods for producing vaccines for respiratory syncytial virus
AR075371A1 (es) 2009-02-05 2011-03-30 Athenix Corp Genes de delta endotoxina variante axmi-r1 y metodos de uso de los mismos
WO2010089302A1 (en) 2009-02-06 2010-08-12 University Of Chile Protein and dna sequence encoding a cold adapted xylanase
WO2010091221A1 (en) 2009-02-06 2010-08-12 Novozymes A/S Polypeptides having alpha-amylase activity and polynucleotides encoding same
SG2014012918A (en) 2009-02-11 2014-04-28 Novozymes Biopharma Dk As Albumin variants and conjugates
DK2398893T3 (en) 2009-02-19 2015-03-23 Novozymes As PROCESS FOR BREWING FOR USING fungal or Bacterial
MY189620A (en) * 2009-02-20 2022-02-21 Malaysian Palm Oil Board A constitutive promoter from oil palm
CA2753287A1 (en) 2009-02-24 2010-09-02 Glaxo Group Limited Antigen-binding constructs
CA2753332A1 (en) 2009-02-24 2010-09-02 Glaxo Group Limited Antigen-binding constructs
CA2753263A1 (en) 2009-02-24 2010-09-02 Glaxo Group Limited Multivalent and/or multispecific rankl-binding constructs
AU2010217663A1 (en) 2009-02-25 2011-09-01 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
EP2401369B1 (en) 2009-02-26 2014-12-24 Codexis, Inc. Beta-glucosidase variant enzymes and related polynucleotides
US20110306139A1 (en) 2009-02-27 2011-12-15 Novozymes A/S Mutant Cells Suitable For Recombinant Polypeptide Production
JP5746055B2 (ja) 2009-02-27 2015-07-08 アテニックス・コーポレーションＡｔｈｅｎｉｘ Ｃｏｒｐｏｒａｔｏｎ 殺虫性タンパク質及びその使用方法
AU2010221183B2 (en) 2009-03-06 2016-02-04 Athenix Corporation Methods and compositions for controlling plant pests
EP2405920A1 (en) 2009-03-06 2012-01-18 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research Novel therapy for anxiety
US20100298211A1 (en) 2009-03-11 2010-11-25 Athenix Corporation Axmi-001, axmi-002, axmi-030, axmi-035, and axmi-045: toxin genes and methods for their use
EP2408908B1 (en) 2009-03-17 2015-05-27 Codexis, Inc. Variant endoglucanases and related polynucleotides
WO2010106068A1 (en) 2009-03-17 2010-09-23 Novozymes A/S Polypeptides having tyrosinase activity and polynucleotides encoding same
WO2010108918A1 (en) 2009-03-24 2010-09-30 Novozymes A/S Polypeptides having acetyl xylan esterase activity and polynucleotides encoding same
WO2010111691A2 (en) 2009-03-27 2010-09-30 Life Technologies Corp Conjugates of biomolecules to nanoparticles
WO2010109053A1 (es) 2009-03-27 2010-09-30 Proyeto De Biomedicina Cima, S.L. Métodos y composiciones para el tratamiento de cirrosis y fibrosis hepática
EP2414506B1 (en) 2009-03-31 2017-05-03 Codexis, Inc. Improved endoglucanases
JP2012522875A (ja) 2009-04-01 2012-09-27 ダニスコ・ユーエス・インク アルファ‐アミラーゼ及びプロテアーゼを含む洗浄組成物及び方法
EP2241323A1 (en) 2009-04-14 2010-10-20 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research Tenascin-W and brain cancers
GB0908770D0 (en) 2009-04-24 2009-07-01 Danisco Method
CN102459614A (zh) 2009-04-29 2012-05-16 巴斯夫植物科学有限公司 具有增强的产量相关性状的植物和用于产生该植物的方法
CA2760266A1 (en) 2009-04-29 2010-11-04 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
MX2011011678A (es) 2009-05-04 2011-12-06 Pioneer Hi Bred Int Mejora de la produccion en plantas mediante la modulacion del factor de transcripcion ap2.
US20120096593A1 (en) 2009-05-06 2012-04-19 Yves Hatzfeld Plants Having Enhanced Yield-Related Traits And/Or Enhanced Abiotic Stress Tolerance And A Method For Making The Same
EP2248893A1 (en) 2009-05-06 2010-11-10 Novozymes A/S DFPase Enzymes from Octopus Vulgaris
MY186558A (en) * 2009-05-13 2021-07-27 Malaysian Palm Oil Board A constitutive promoter from oil palm type 2
CA2761767A1 (en) 2009-05-19 2010-11-25 Danisco A/S Use of amylase and lipolytic enzyme in bread
MX355638B (es) 2009-05-21 2018-03-26 Verenium Corp Fitasas, ácidos nucléicos que las codifican, y métodos de hacerlas y usarlas.
WO2010135695A2 (en) 2009-05-22 2010-11-25 Curna, Inc. TREATMENT OF TRANSCRIPTION FACTOR E3 (TFE3) and INSULIN RECEPTOR SUBSTRATE 2 (IRS2) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO TFE3
US9989533B2 (en) 2009-05-28 2018-06-05 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Anti-TNF induced apoptosis (ATIA) diagnostic markers and therapies
DK2435568T3 (da) 2009-05-29 2014-09-08 Morphosys Ag Samling af syntetiske antistoffer til behandling af sygdom
US20100304437A1 (en) 2009-05-29 2010-12-02 Novozymes, Inc. Methods for enhancing the degradation or conversion of cellulosic material
US8278507B2 (en) 2009-06-02 2012-10-02 Novozymes, Inc. Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
WO2010147825A1 (en) 2009-06-09 2010-12-23 Pioneer Hi-Bred International, Inc. Early endosperm promoter and methods of use
AR077042A1 (es) 2009-06-12 2011-07-27 Danisco Metodo para tratar una composicion que contiene pirofiofitina
US20100317539A1 (en) * 2009-06-12 2010-12-16 Guo-Liang Yu Library of Engineered-Antibody Producing Cells
CA2762616C (en) 2009-06-16 2015-08-18 Codexis, Inc. Beta-glucosidase variant enzymes and related polynucleotides
EA201270035A1 (ru) 2009-06-19 2012-09-28 Басф Плант Сайенс Компани Гмбх Растения с улучшенными характеристиками урожайности и способ их получения
BRPI1015234A2 (pt) 2009-06-22 2018-02-20 Medimmune Llc regiões fc projetadas para conjugação sítio específica.
HUE055413T2 (hu) 2009-06-22 2021-11-29 Codexis Inc Ketoreduktáz-közvetített sztereoszelektív útvonal alfa-klóralkoholokhoz
CN102712919B (zh) 2009-06-26 2017-06-23 诺维信北美公司 热稳定性碳酸酐酶及其用途
CN102648281B (zh) 2009-07-02 2017-04-05 阿森尼克斯公司 Axmi‑205杀虫基因和它的使用方法
WO2011000924A1 (en) 2009-07-03 2011-01-06 Abbott Products Gmbh Spray-dried amylase, pharmaceutical preparations comprising the same and use
WO2011005823A1 (en) 2009-07-07 2011-01-13 Castle Linda A Crystal structure of glyphosate acetyltransferase (glyat) and methods of use
DK2451957T3 (en) 2009-07-07 2018-02-05 Novozymes Inc POLYPEPTIDES WITH CELLULOLYSE ENHANCING ACTIVITY AND POLYNUCLEOTIDES CODING THEM
US8614081B2 (en) 2009-07-23 2013-12-24 Codexis, Inc. Nitrilase biocatalysts
WO2011009747A1 (en) 2009-07-24 2011-01-27 Novozymes A/S Carbohydrate oxidases
WO2011009182A2 (pt) 2009-07-24 2011-01-27 Embrapa - Empresa Brasileira De Pesquisa Agropecuária Molécula de ácido nucléico isolada, construção gênica, vetor, célula transgênica, método para obtenção de uma célula e de uma planta transgênica, polipeptídeo isolado e purificado, composição pesticida biodegradável, método para o controle de uma praga, método de obtenção de linhagens transgênicas resistentes a um inseto praga
WO2011014458A1 (en) 2009-07-28 2011-02-03 Novozymes, Inc. Polypeptides having phytase activity and polynucleotides encoding same
MX2012001426A (es) 2009-07-31 2012-03-26 Athenix Corp Familia de genes plaguicidas axmi-192 y metodos para su uso.
WO2011018421A1 (en) 2009-08-10 2011-02-17 Morphosys Ag Novel screening strategies for the identification of binders
WO2011020024A2 (en) 2009-08-13 2011-02-17 The Johns Hopkins University Methods of modulating immune function
ES2553440T3 (es) * 2009-08-13 2015-12-09 Crucell Holland B.V. Anticuerpos contra el virus sincitial respiratorio humano (VSR) y método de uso
EP2467473B1 (en) 2009-08-19 2016-03-23 Codexis, Inc. Ketoreductase polypeptides for the preparation of phenylephrine
US20120144529A1 (en) 2009-08-19 2012-06-07 Basf Plant Science Company Gmbh Plants Having Enhanced Yield-Related Traits and a Method for Making the Same
MX2012002113A (es) 2009-08-20 2012-08-08 Pioneer Hi Bred Int Expresion funcional del transportador de nitrato de levadura entremezclado (ynti) en el maiz para mejorar la captacion de nitrato bajo ambiente de bajo contenido de nitrato.
CN102482658B (zh) 2009-08-21 2015-07-15 诺维信公司 具有异淀粉酶活性的多肽和编码该多肽的多核苷酸
EP2292266A1 (en) 2009-08-27 2011-03-09 Novartis Forschungsstiftung, Zweigniederlassung Treating cancer by modulating copine III
WO2011028643A1 (en) 2009-09-01 2011-03-10 Novozymes, Inc. Methods for improving malic acid production in filamentous fungi
EP2473604B1 (en) 2009-09-04 2017-01-11 Codexis, Inc. Variant cbh2 cellulases and related polynucleotides
JP2013503647A (ja) 2009-09-09 2013-02-04 ブラスケム ソシエダッド アノニマ ｎ−プロパノールを製造するための微生物および方法
US8293483B2 (en) 2009-09-11 2012-10-23 Epitomics, Inc. Method for identifying lineage-related antibodies
EP3269804B1 (en) 2009-09-17 2020-11-11 Novozymes, Inc. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
HUE037123T2 (hu) 2009-09-18 2018-08-28 Codexis Inc Csökkentett kodon mutagenezis
US20110189183A1 (en) 2009-09-18 2011-08-04 Robert Anthony Williamson Antibodies against candida, collections thereof and methods of use
WO2011035029A1 (en) 2009-09-18 2011-03-24 Novozymes, Inc. Polypeptides having beta-glucosidase activity and polynucleotides encoding same
US20120244170A1 (en) 2009-09-22 2012-09-27 Rafal Ciosk Treating cancer by modulating mex-3
BR112012006487A8 (pt) 2009-09-25 2018-04-24 Novozymes As variante de uma subtilisina precursora, polinucleotídeo isolado, construção de ácido nucleico, vetor de expressão, célula hospedeira, composição de limpeza ou detergente, e, método para produzir uma variante
US20120252106A1 (en) 2009-09-25 2012-10-04 Novozymes A/S Use of Protease Variants
CA2772983A1 (en) 2009-09-25 2011-03-31 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
KR101754083B1 (ko) 2009-09-25 2017-07-05 바스프 플랜트 사이언스 컴퍼니 게엠베하 향상된 수확량 관련 형질을 갖는 식물 및 이의 제조 방법
CN107338233A (zh) 2009-09-29 2017-11-10 诺维信股份有限公司 具有木聚糖酶活性的多肽和编码该多肽的多核苷酸
ES2560805T3 (es) 2009-09-29 2016-02-22 Novozymes Inc. Polipéptidos con actividad mejoradora celulolítica y polinucleótidos que los codifican
CA2775244A1 (en) 2009-09-30 2011-04-07 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
CN102695720B (zh) 2009-09-30 2018-02-16 诺维信股份有限公司 具有纤维素分解增强活性的多肽和编码该多肽的多核苷酸
US8222012B2 (en) * 2009-10-01 2012-07-17 E. I. Du Pont De Nemours And Company Perhydrolase for enzymatic peracid production
US9243262B2 (en) 2009-10-02 2016-01-26 Syngenta Participations Ag Insecticidal proteins
LT2486141T (lt) 2009-10-07 2018-05-25 Macrogenics, Inc. Fc regioną turintys polipeptidai, pasižymintys pagerinta efektorine funkcija dėl fukozilinimo laipsnio pasikeitimų, ir jų naudojimo būdai
WO2011045352A2 (en) 2009-10-15 2011-04-21 Novartis Forschungsstiftung Spleen tyrosine kinase and brain cancers
UA109884C2 (uk) 2009-10-16 2015-10-26 Поліпептид, що має активність ферменту фосфатидилінозитол-специфічної фосфоліпази с, нуклеїнова кислота, що його кодує, та спосіб його виробництва і застосування
UA111708C2 (uk) 2009-10-16 2016-06-10 Бандж Ойлз, Інк. Спосіб рафінування олії
DE112010003738T5 (de) 2009-10-22 2013-08-14 Basf Plant Science Company Gmbh Pflanzen mit gesteigerten Ertragsmerkmalen und Verfahren zu deren Herstellung
US8541651B2 (en) 2009-10-23 2013-09-24 Novozymes, Inc. Cellobiohydrolase variants and polynucleotides encoding same
IN2012DN03073A (zh) 2009-10-26 2015-07-31 Pioneer Hi Bred Int
CN102666847B (zh) 2009-10-29 2015-12-09 诺维信股份有限公司 具有纤维二糖水解酶活性的多肽和编码该多肽的多核苷酸
WO2011051327A2 (en) 2009-10-30 2011-05-05 Novartis Ag Small antibody-like single chain proteins
RU2607374C2 (ru) 2009-10-30 2017-01-10 Новозаймс Байофарма Дк А/С Варианты альбумина
US20120213801A1 (en) 2009-10-30 2012-08-23 Ekaterina Gresko Phosphorylated Twist1 and cancer
WO2011051466A1 (en) 2009-11-02 2011-05-05 Novartis Ag Anti-idiotypic fibronectin-based binding molecules and uses thereof
US9267148B2 (en) 2009-11-02 2016-02-23 Plant Sensory Systems, Llc Methods for the biosynthesis of taurine or hypotaurine in cells
US10207240B2 (en) 2009-11-03 2019-02-19 Gen9, Inc. Methods and microfluidic devices for the manipulation of droplets in high fidelity polynucleotide assembly
CN103068976A (zh) 2009-11-06 2013-04-24 诺维信股份有限公司 具有纤维二糖水解酶活性的多肽和编码该多肽的多核苷酸
CN102639697B (zh) 2009-11-06 2015-03-25 诺维信股份有限公司 具有木聚糖酶活性的多肽和编码该多肽的多核苷酸
AU2010318024A1 (en) 2009-11-13 2012-06-21 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
GB0920089D0 (en) 2009-11-17 2009-12-30 Danisco Method
WO2011063198A2 (en) 2009-11-20 2011-05-26 St. Jude Children's Research Hospital Methods and compositions for modulating the activity of the interleukin-35 receptor complex
BR112012012374A2 (pt) 2009-11-23 2015-09-15 Du Pont semente de planta com maior teor de óleo, planta, método para aumentar ainda mais o teor de óleo em uma semente de planta de alto teor de óleo, método par avaliar o teor de óleo aumentado em uma semente de planta, polinucleotídeo isolado polipeptídeo isolado
WO2011066185A1 (en) 2009-11-25 2011-06-03 Gen9, Inc. Microfluidic devices and methods for gene synthesis
MX2012006016A (es) 2009-11-25 2012-06-25 Codexis Inc Variantes de beta-glucosidasa recombinante para produccion de azucares solubles de biomasa celulosica.
BR112012012192A2 (pt) 2009-11-25 2017-07-04 Codexis Inc b-glicosidades variantes recombinantes thermoascus aurantiacus para produção de açúcares fermentáveis a partir da biomassa celulósica
EP3085791A1 (en) 2009-11-25 2016-10-26 Gen9, Inc. Methods and apparatuses for chip-based dna error reduction
JP2013511979A (ja) 2009-11-27 2013-04-11 ビーエーエスエフ プラント サイエンス カンパニー ゲーエムベーハー キメラエンドヌクレアーゼおよびその使用
EP2504429B1 (en) 2009-11-27 2016-04-20 BASF Plant Science Company GmbH Chimeric endonucleases and uses thereof
CA2782014C (en) 2009-11-27 2021-08-31 Basf Plant Science Company Gmbh Optimized endonucleases and uses thereof
CA2782128C (en) 2009-11-30 2019-01-15 Novozymes A/S Polypeptides having glucoamylase activity and polynucleotides encoding same
CA2782154C (en) 2009-11-30 2018-10-16 Novozymes A/S Polypeptides having glucoamylase activity and polynucleotides encoding same
CN104169417B (zh) 2009-12-01 2020-05-08 诺维信公司 具有葡糖淀粉酶活性的多肽及其编码多核苷酸
TW201125577A (en) 2009-12-02 2011-08-01 Novozymes As Use of defensins for treatment of infective endocarditis
MX2012006397A (es) 2009-12-02 2012-11-30 Amgen Inc PROTEINAS DE ENLACE QUE ENLAZAN A FGFR1C HUMANO, ß-KLOTHO HUMANA Y TANTO FGFR1C HUMANO COMO ß-KLOTHO HUMANA.
US20130023028A1 (en) 2009-12-03 2013-01-24 Novozymes South Asia Pvt. Ltd. Variants Of A Polypeptide With Lipolytic Activity and Improved Stability
SI2510089T1 (sl) 2009-12-08 2015-12-31 Codexis, Inc. Sinteza prazolnih spojin
WO2011070101A1 (en) 2009-12-09 2011-06-16 Novozymes A/S Methods of producing gh8 xylanase variants
DK2510093T3 (en) 2009-12-11 2018-06-06 Novozymes North America Inc protease Variants
US8653024B2 (en) 2009-12-11 2014-02-18 Adenium Biotech Aps Use of AMPs for treatment of UTI/cystitis
EP2501792A2 (en) 2009-12-29 2012-09-26 Novozymes A/S Gh61 polypeptides having detergency enhancing effect
CA2786001A1 (en) 2009-12-31 2011-07-07 Pioneer Hi-Bred International, Inc. Engineering plant resistance to diseases caused by pathogens
DK2521774T3 (en) 2010-01-04 2016-09-26 Novozymes As ALFA AMYLASE VARANTS AND POLYNUCLEOTIDES CODING THESE
US9217144B2 (en) 2010-01-07 2015-12-22 Gen9, Inc. Assembly of high fidelity polynucleotides
US20120295793A1 (en) 2010-01-22 2012-11-22 Proteus Methods of generating modified polynucleotide libraries and methods of using the same for directed protein evolution
EP2529019B1 (en) 2010-01-25 2014-10-01 Bayer CropScience NV Methods for manufacturing plant cell walls comprising chitin
WO2011092233A1 (en) 2010-01-29 2011-08-04 Novartis Ag Yeast mating to produce high-affinity combinations of fibronectin-based binders
CA2789629A1 (en) 2010-02-10 2011-08-18 Immunogen, Inc. Cd20 antibodies and uses thereof
EP2357220A1 (en) 2010-02-10 2011-08-17 The Procter & Gamble Company Cleaning composition comprising amylase variants with high stability in the presence of a chelating agent
WO2011100265A2 (en) 2010-02-10 2011-08-18 Codexis, Inc. Processes using amino acid dehydrogenases and ketoreductase-based cofactor regenerating system
CN113186178A (zh) 2010-02-10 2021-07-30 诺维信公司 在螯合剂存在下具有高稳定性的变体和包含变体的组合物
AU2011218131B2 (en) 2010-02-18 2016-03-17 BASF Agricultural Solutions Seed US LLC AXMI221z, AXMI222z, AXMI223z, AXMI224z, and AXMI225z delta-endotoxin genes and methods for their use
CN102892886A (zh) 2010-02-18 2013-01-23 阿森尼克斯公司 AXMI218、AXMI219、AXMI220、AXMI226、AXMI227、AXMI228、AXMI229、AXMI230、和AXMI231δ-内毒素基因以及它们的使用方法
US9388423B2 (en) 2010-02-24 2016-07-12 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
AU2011219927A1 (en) 2010-02-24 2012-08-23 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and a method for making the same
WO2011104284A1 (en) 2010-02-25 2011-09-01 Novozymes A/S Polypeptides having antimicrobial activity
WO2011104339A1 (en) 2010-02-25 2011-09-01 Novozymes A/S Variants of a lysozyme and polynucleotides encoding same
EP2361985A1 (en) 2010-02-26 2011-08-31 BASF Plant Science Company GmbH Plants having enhanced yield-related traits and a method for making the same
EP2361927A1 (en) 2010-02-26 2011-08-31 BASF Plant Science Company GmbH Plants having enhanced yield-related traits and a method for making the same
CN102884182A (zh) 2010-03-03 2013-01-16 诺维信股份有限公司 木聚糖酶变体及编码其的多核苷酸
US20130004519A1 (en) 2010-03-05 2013-01-03 Ruth Chiquet-Ehrismann Smoci, tenascin-c and brain cancers
EP2545150B1 (en) 2010-03-12 2018-07-25 DuPont Nutrition Biosciences ApS Process
AU2011228707A1 (en) 2010-03-18 2012-09-27 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and method for making the same
WO2011114251A1 (en) 2010-03-18 2011-09-22 Danisco A/S Foodstuff
MX2012010600A (es) 2010-03-18 2012-10-05 Basf Plant Science Co Gmbh Plantas que tienen mejores rasgos relacionados con el redimiento y un metodo para producirlas.
US9556248B2 (en) 2010-03-19 2017-01-31 The Board Of Trustees Of The Leland Stanford Junior University Hepatocyte growth factor fragments that function as potent met receptor agonists and antagonists
EA201290936A1 (ru) 2010-03-19 2013-05-30 Басф Плант Сайенс Компани Гмбх Растения с улучшенными характеристиками урожайности и способ их получения
WO2011114312A1 (en) 2010-03-19 2011-09-22 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and method for making same
EP2371845A1 (en) 2010-03-22 2011-10-05 BASF Plant Science Company GmbH Plants having enhanced yield-related traits and a method for making the same
AU2011231563B2 (en) 2010-03-26 2015-01-29 Novozymes A/S Thermostable phytase variants
EP2553109B1 (en) 2010-03-28 2018-10-24 Novozymes A/S Regioselective hydroxylation of aliphatic hydrocarbons employing a fungal peroxygenase
ES2636216T3 (es) 2010-03-30 2017-10-05 Novozymes North America, Inc. Procesos de producción de un producto de fermentación
EP2553108A4 (en) 2010-03-31 2015-01-28 Codexis Inc PRODUCTION OF MONOTERPENES
WO2011123450A1 (en) 2010-03-31 2011-10-06 Novozymes, Inc. Cellobiohydrolase variants and polynucleotides encoding same
CN102939304B (zh) 2010-04-09 2017-04-19 阿尔布麦狄克斯公司 白蛋白衍生物和变体
DK2558484T3 (en) 2010-04-14 2016-03-29 Novozymes As Polypeptides having glucoamylase activity and polynucleotides encoding them
US8383793B2 (en) 2010-04-15 2013-02-26 St. Jude Children's Research Hospital Methods and compositions for the diagnosis and treatment of cancer resistant to anaplastic lymphoma kinase (ALK) kinase inhibitors
LT2558577T (lt) 2010-04-16 2019-03-12 Nuevolution A/S Bifunkciniai kompleksai ir tokių kompleksų gamybos ir naudojimo būdai
US20130034543A1 (en) 2010-04-19 2013-02-07 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Resear Modulating xrn1
HUE026367T2 (en) 2010-05-04 2016-06-28 Codexis Inc Biocatalysts of ezetimibe synthesis
EP2566894A1 (en) 2010-05-06 2013-03-13 Novartis AG Compositions and methods of use for therapeutic low density lipoprotein - related protein 6 (lrp6) multivalent antibodies
US20110277182A1 (en) 2010-05-06 2011-11-10 E.I. Dupont De Nemours And Company Maize acc synthase 3 gene and protein and uses thereof
NZ603829A (en) 2010-05-06 2015-03-27 Novartis Ag Compositions and methods of use for therapeutic low density lipoprotein -related protein 6 (lrp6) antibodies
JP2013528374A (ja) 2010-05-10 2013-07-11 パーシード セラピューティクス リミテッド ライアビリティ カンパニー Ｖｌａ４のポリペプチド阻害剤
PT2569010T (pt) 2010-05-14 2017-05-31 Amgen Inc Formulações de anticorpos de concentração elevada
CN103429265A (zh) 2010-05-25 2013-12-04 国家医疗保健研究所 用于治疗和预防hcv感染的抗包膜抗体和抗受体抗体组合物
US20110294170A1 (en) 2010-05-28 2011-12-01 Codexis, Inc. Pentose Fermentation By a Recombinant Microorganism
EP2576606B1 (en) 2010-06-04 2014-12-10 Novozymes, Inc. C4 dicarboxylic acid production in filamentous fungi
WO2011154485A1 (en) 2010-06-10 2011-12-15 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research Treating cancer by modulating mammalian sterile 20-like kinase 3
US20130084358A1 (en) 2010-06-11 2013-04-04 Novozymes A/S Method To Reduce Biogenic Amine Content in Food
US8835604B2 (en) 2010-06-12 2014-09-16 Adenium Biotech Aos Antimicrobial peptide variants and polynucleotides encoding same
BR112012032176A2 (pt) 2010-06-17 2015-09-08 Dupont Nutrition Biosci Aps processo de tratamento de sementes que contém óleo, método de obtenção de óleo de sementes vegetais, processo de produção de um óleo vegetal refinado e óleo vegetal bruto ou refinado que pode ser obtido por meio de um método ou processo
WO2011159910A2 (en) 2010-06-17 2011-12-22 Codexis, Inc. Biocatalysts and methods for the synthesis of (s)-3-(1-aminoethyl)-phenol
BR112012029891A2 (pt) 2010-06-21 2021-07-06 Novozymes Inc polipeptídeo isolado, uso do mesmo, composição, polinucleotídeo isolado, construção de ácido nucleico ou vetor de expressão, célula hospedeira, e, métodos para produzir e para aumentar a produção de um ácido dicarboxílico c4.
WO2011163270A1 (en) 2010-06-21 2011-12-29 Novozymes, Inc. Methods for improved c4-dicarboxylic acid production in filamentous fungi
CN105063063A (zh) 2010-06-24 2015-11-18 巴斯夫植物科学有限公司 具有增强的产量相关性状的植物和用于制备该植物的方法
WO2011163590A1 (en) 2010-06-25 2011-12-29 E. I. Du Pont De Nemours And Company Compositions and methods for enhancing resistance to northern leaf blight in maize
KR101429475B1 (ko) 2010-06-25 2014-08-22 바스프 플랜트 사이언스 컴퍼니 게엠베하 향상된 수확량 관련 형질을 갖는 식물 및 이의 제조 방법
CA2804019A1 (en) 2010-06-28 2012-01-12 Codexis, Inc. Fatty alcohol forming acyl reductases (fars) and methods of use thereof
AU2011272878B2 (en) 2010-06-30 2015-04-23 Codexis, Inc. Highly stable beta-class carbonic anhydrases useful in carbon capture systems
WO2012003379A1 (en) 2010-06-30 2012-01-05 Novozymes A/S Polypeptides having beta-glucosidase activity and polynucleotides encoding same
GB201011513D0 (en) 2010-07-08 2010-08-25 Danisco Method
AR082149A1 (es) 2010-07-09 2012-11-14 Calmune Corp Anticuerpos contra el virus sincicial respiratorio (rsv) humano y metodos para su uso
RU2016149485A (ru) 2010-07-12 2018-10-31 Селджин Корпорейшн Твердые формы ромидепсина и их применение
DK3741883T3 (da) 2010-07-16 2023-02-20 Adimab Llc Antistofbiblioteker
AU2011277925A1 (en) 2010-07-16 2013-02-21 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and method for making the same
WO2012016245A2 (en) 2010-07-30 2012-02-02 Novartis Ag Fibronectin cradle molecules and libraries thereof
EP2598629B1 (en) 2010-07-30 2020-04-08 Cleanvantage LLC Aspergillus containing beta-glucosidase, beta-glucosidases and nucleic acids encoding the same
SG187682A1 (en) 2010-08-02 2013-03-28 Macrogenics Inc Covalent diabodies and uses thereof
EP2603594B1 (en) 2010-08-12 2018-04-25 Novozymes, Inc. Compositions comprising a polypeptide having cellulolytic enhancing activity and a bicyclic compound and uses thereof
AU2011289286B2 (en) 2010-08-13 2015-02-12 Pioneer Hi-Bred International, Inc. Methods and compositions for targeting sequences of interest to the chloroplast
WO2012022734A2 (en) 2010-08-16 2012-02-23 Medimmune Limited Anti-icam-1 antibodies and methods of use
CA2808098C (en) 2010-08-16 2020-03-10 Merck Sharp & Dohme Corp. Process for preparing aminocyclohexyl ether compounds
WO2012024662A2 (en) 2010-08-20 2012-02-23 Codexis, Inc. Expression constructs comprising fungal promoters
CN103080306B (zh) 2010-08-20 2015-02-18 科德克希思公司 糖苷水解酶61家族蛋白在纤维素加工中的用途
MY162825A (en) 2010-08-20 2017-07-31 Novartis Ag Antibodies for epidermal growth factor receptor 3 (her3)
CA2807785A1 (en) 2010-08-23 2012-03-01 Pioneer Hi-Bred International, Inc. Novel defensin variants and methods of use
EP2609196B2 (en) 2010-08-24 2020-05-06 Novozymes A/S Heat-stable persephonella carbonic anhydrases and their use
AU2011294767A1 (en) 2010-08-24 2013-03-21 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and method for making the same
JP5963749B2 (ja) 2010-08-26 2016-08-03 イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニーＥ．Ｉ．Ｄｕ Ｐｏｎｔ Ｄｅ Ｎｅｍｏｕｒｓ Ａｎｄ Ｃｏｍｐａｎｙ 変異δ９エロンガーゼおよび多価不飽和脂肪酸の製造におけるそれらの使用
CN103080317B (zh) 2010-08-26 2017-07-11 纳幕尔杜邦公司 突变型hpgg基序和hdash基序δ‑5去饱和酶以及它们在制备多不饱和脂肪酸中的用途
US8629325B2 (en) 2010-08-30 2014-01-14 Novozymes A/S Polypeptides having beta-glucosidase activity and polynucleotides encoding same
EP2735611B1 (en) 2010-08-30 2018-11-21 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
WO2012030811A1 (en) 2010-08-30 2012-03-08 Novozymes A/S Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
US20130212746A1 (en) 2010-08-30 2013-08-15 Novoyzmes A/S Polypeptides Having Hemicellulolytic Activity And Polynucleotides Encoding Same
WO2012030849A1 (en) 2010-08-30 2012-03-08 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
WO2012030844A1 (en) 2010-08-30 2012-03-08 Novozymes A/S Polypeptides having endoglucanase activity and polynucleotides encoding same
EP2614080A1 (en) 2010-09-10 2013-07-17 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research Phosphorylated twist1 and metastasis
US8859502B2 (en) 2010-09-13 2014-10-14 Celgene Corporation Therapy for MLL-rearranged leukemia
WO2012035103A1 (en) 2010-09-16 2012-03-22 Novozymes A/S Lysozymes
MX2013003159A (es) 2010-09-22 2013-05-01 Bayer Ip Gmbh Uso de agentes de control biologico o quimico para controlar insectos y nematodos en cultivos resistentes.
CN103237891B (zh) 2010-09-30 2017-07-14 诺维信股份有限公司 具有纤维素分解增强活性的多肽变体及其编码多核苷酸
MX2013003236A (es) 2010-09-30 2013-05-30 Novozymes Inc Variantes de polipeptidos que tienen actividad potenciadora celulolitica y polinucleotidos que codifican a los mismos.
US9688975B2 (en) 2010-10-01 2017-06-27 Novozymes, Inc. Beta-glucosidase variants and polynucleotides encoding same
JP5944908B2 (ja) 2010-10-01 2016-07-05 ノボザイムス アクティーゼルスカブ エンドペプチダーゼ活性を有するポリペプチド及びそれをコードするポリヌクレオチド
WO2012051055A2 (en) 2010-10-06 2012-04-19 Bp Corporation North America Inc. Variant cbh i polypeptides
US20130260423A1 (en) 2010-10-26 2013-10-03 Novozymes North America, Inc. Methods of Saccharifying Sugar Cane Trash
US20130280775A1 (en) 2010-10-29 2013-10-24 Novozymes, Inc. Recombinant N-propanol and Isopropanol Production
CN103282376A (zh) 2010-10-29 2013-09-04 诺维信公司 具有琥珀酰CoA:乙酰乙酸转移酶活性的多肽及其编码多核苷酸
DK2635689T3 (en) 2010-11-02 2015-07-06 Novozymes Inc METHODS OF PREPARATION OF cellulosic material WITH A GH61-POLYPEPTIDE
EP2635594B1 (en) 2010-11-04 2017-01-11 Novozymes Inc. Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
KR101973930B1 (ko) 2010-11-05 2019-04-29 자임워크스 인코포레이티드 Fc 도메인 내의 돌연변이를 갖는 안정한 이종이량체 항체 디자인
ES2649555T3 (es) 2010-11-08 2018-01-12 Novozymes A/S Polipéptidos con actividad glucoamilasa y polinucleótidos que los codifican
CA2815804A1 (en) 2010-11-08 2012-05-18 Novozymes A/S Polypeptides having glucoamylase activity and polynucleotides encoding same
EP2638166A4 (en) 2010-11-10 2014-04-30 Basf Plant Science Co Gmbh PLANTS HAVING IMPROVED YIELD-RELATED ASPECTS AND METHOD FOR OBTAINING SAME
MX356647B (es) 2010-11-12 2018-06-07 Novozymes As Polipeptidos que tienen actividad de fosfolipasa c y polinucleotidos que codifican los mismos.
EP2638153B1 (en) 2010-11-12 2017-07-12 Novozymes Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
US20140093506A1 (en) 2010-11-15 2014-04-03 Marc Buehler Anti-fungal-agents
MX2013004758A (es) 2010-11-18 2013-06-28 Novozymes Inc Polipeptidos quimericos que tienen actividad de incremento celulolitico y polinucleotidos que codifican para los mismos.
ES2696548T3 (es) 2010-11-19 2019-01-16 Morphosys Ag Una colección de secuencias de anticuerpos y su uso
BR112013013003A2 (pt) 2010-11-24 2016-08-09 Glaxo Group Ltd proteína de ligação de antígeno, e, composição farmacêutica
US20130245233A1 (en) 2010-11-24 2013-09-19 Ming Lei Multispecific Molecules
WO2012078656A1 (en) 2010-12-06 2012-06-14 Novozymes North America, Inc. Methods of hydrolyzing oligomers in hemicellulosic liquor
WO2012078800A2 (en) 2010-12-08 2012-06-14 Codexis, Inc. Biocatalysts and methods for the synthesis of armodafinil
CN103270160B (zh) 2010-12-22 2015-09-09 先锋国际良种公司 病毒启动子、其截短物以及使用方法
MX2013007087A (es) 2010-12-22 2013-07-29 Du Pont Promotor viral, truncamientos de este y metodos de uso.
BR112013015515A2 (pt) 2010-12-28 2018-04-24 Pioneer Hi Bred Int molécula de ácido nucleico isolada, construto de dna, célula hospedeira, planta transgênica, semente transformada da planta, polipeptídeo isolado com atividade pesticida, composição, método para controlar uma população de praga de lepidóptero, método para matar uma praga de lepidóptero, método para produzir um polipeptídeo com atividade pesticida, planta que tem incorporado de maneira estável em seu genoma um construto de dna, método para proteger uma planta contra uma praga
BR112013017955A2 (pt) 2011-01-14 2019-09-24 Florida Res Foundation Inc métodos para a preparação de uma planta cítrica com resistência intensificada ao cancro cítrico e para produzir fruta cítrica, molécula de ácido nucleico, cassete de expressão ou vetor, planta ou célula de planta, molécula de ácido nucleico, e, célula hospedeira
EP2665819A4 (en) 2011-01-20 2014-09-10 Basf Plant Science Co Gmbh PLANTS HAVING AMPLIFIED YIELD CHARACTERS AND PROCESS FOR PRODUCTION THEREOF
BR112013019247B1 (pt) 2011-01-26 2021-03-30 Novozymes A/S Método para produzir um polipeptídeo
MX2013007720A (es) 2011-01-26 2013-08-09 Novozymes As Polipeptidos que tienen actividad celobiohidrolasa y polinucleotidos que codifican para los mismos.
MX337942B (es) 2011-01-26 2016-03-29 Novozymes As Polipeptidos que tienen actividad de endoglucanasa y polinucleotidos que codifican los mismos.
EP2668266B1 (en) 2011-01-26 2018-01-03 Novozymes A/S Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
CN103517986B (zh) 2011-01-26 2016-12-07 诺维信公司 具有纤维二糖水解酶活性的多肽及编码该多肽的多核苷酸
US20120189633A1 (en) 2011-01-26 2012-07-26 Kolltan Pharmaceuticals, Inc. Anti-kit antibodies and uses thereof
CN103459605B (zh) 2011-01-31 2016-08-24 诺维信北美公司 用于酶法精制经预处理的纤维素材料以供糖化的工艺
WO2012106579A1 (en) 2011-02-03 2012-08-09 Bristol-Myers Squibb Company Amino acid dehydrogenase and its use in preparing amino acids from keto acids
WO2012112411A1 (en) 2011-02-15 2012-08-23 Pioneer Hi-Bred International, Inc. Root-preferred promoter and methods of use
CN103476915A (zh) 2011-02-16 2013-12-25 诺维信公司 包含金属蛋白酶的去污剂组合物
WO2012110564A1 (en) 2011-02-16 2012-08-23 Novozymes A/S Detergent compositions comprising m7 or m35 metalloproteases
MX2013009176A (es) 2011-02-16 2013-08-29 Novozymes As Composiciones detergentes que comprenden metaloproteasas.
WO2012114232A1 (en) 2011-02-23 2012-08-30 Dupont Nutrition Biosciences Aps Process
EP2678352B1 (en) 2011-02-23 2017-12-06 Novozymes, Inc. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
US9493749B2 (en) 2011-02-23 2016-11-15 Dupont Nutrition Biosciences Aps Process modifications to enhance chlorophyll degradation
AR085253A1 (es) 2011-02-23 2013-09-18 Danisco Metodo para producir una clorofilasa
CN103492551A (zh) 2011-02-28 2014-01-01 诺维信股份有限公司 用于生产c4-二羧酸的微生物
WO2012117324A1 (en) 2011-02-28 2012-09-07 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and producing methods thereof
CA2826859A1 (en) 2011-02-28 2012-09-07 Basf Plant Science Company Gmbh Plants having enhanced yield-related traits and producing methods thereof
EP2681322A4 (en) 2011-03-01 2015-05-06 Basf Plant Science Co Gmbh PLANTS HAVING IMPROVED YIELD CHARACTERS AND PROCESS FOR PRODUCING SAME
EP2681242B1 (en) 2011-03-01 2018-01-24 Amgen Inc. Sclerostin and dkk-1 bispecific binding agents
WO2012122518A1 (en) 2011-03-09 2012-09-13 Novozymes A/S Methods of increasing the cellulolytic enhancing activity of a polypeptide
US8878007B2 (en) 2011-03-10 2014-11-04 Pioneer Hi Bred International Inc Bacillus thuringiensis gene with lepidopteran activity
US9409958B2 (en) 2011-03-10 2016-08-09 Novozymes, Inc. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
US9428558B2 (en) 2011-03-23 2016-08-30 Novozymes A/S Methods for producing secreted polypeptides
WO2012127002A1 (en) 2011-03-23 2012-09-27 Novozymes A/S Sweet-tasting polypeptide from gram-positive bacteria
EP3333258A3 (en) 2011-03-25 2018-07-25 Novozymes A/S Method for degrading or converting cellulosic material
EA029956B1 (ru) 2011-03-25 2018-06-29 Эмджен Инк. Кристаллы антител против склеростина и составы на их основе
US9695440B2 (en) 2011-03-30 2017-07-04 Athenix Corp. AXMI232, AXMI233, and AXMI249 toxin genes and methods for their use
US9321814B2 (en) 2011-03-30 2016-04-26 Athenix Corp. AXMI238 toxin gene and methods for its use
US8663962B2 (en) 2011-03-30 2014-03-04 Codexis, Inc. Pentose fermentation by a recombinant microorganism
WO2012135659A2 (en) 2011-03-31 2012-10-04 Novozymes A/S Methods for enhancing the degradation or conversion of cellulosic material
US20140080182A1 (en) 2011-03-31 2014-03-20 Novozymes, Inc. Cellulose Binding Domain Variants and Polynucleotides Encoding Same
MX344229B (es) 2011-04-05 2016-12-07 Athenix Corp Gen insecticida variante axmi115 y metodos para utilizarlo.
BR112013025811A2 (pt) 2011-04-08 2016-11-29 Danisco Us Inc "composição e método para remover uma mancha de base lipídica de uma superfície"
WO2012142302A2 (en) 2011-04-13 2012-10-18 Codexis, Inc. Biocatalytic process for preparing eslicarbazepine and analogs thereof
US20140157443A1 (en) 2011-04-14 2014-06-05 St. Jude Children's Research Hospital Methods and compositions for detecting and modulating a novel mtor complex
US9034322B2 (en) 2011-04-21 2015-05-19 The Rockefeller University Streptococcus bacteriophage lysins for detection and treatment of gram positive bacteria
US9926547B2 (en) 2011-04-28 2018-03-27 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
DK2702162T3 (da) 2011-04-29 2020-05-18 Novozymes Inc Fremgangsmåder til forbedring af nedbrydningen eller omdannelsen af celluloseholdigt materiale
WO2012150319A1 (en) 2011-05-05 2012-11-08 Novozymes Biopharma Dk A/S Albumin variants
US8993286B2 (en) 2011-05-19 2015-03-31 Novozymes, Inc. Methods for enhancing the degradation of cellulosic material with chitin binding proteins
WO2012159007A1 (en) 2011-05-19 2012-11-22 Novozymes, Inc. Methods for enhancing the degradation of cellulosic material with chitin binding proteins
US9376495B2 (en) 2011-05-21 2016-06-28 Macrogenics, Inc. Deimmunized serum-binding domains and their use in extending serum half-life
EP2527448A1 (en) 2011-05-23 2012-11-28 Novozymes A/S Simultaneous site-specific integrations of multiple gene-copies in filamentous fungi
EP2527432A1 (en) 2011-05-23 2012-11-28 Novozymes A/S Bi-directional cytosine deaminase-encoding selection marker
US9150625B2 (en) 2011-05-23 2015-10-06 E I Du Pont De Nemours And Company Chloroplast transit peptides and methods of their use
CA2837169C (en) 2011-05-24 2021-11-09 Zyngenia, Inc. Multispecific complexes comprising angiopoietin-2-binding peptide and their uses
EP2717911A1 (en) 2011-06-06 2014-04-16 Novartis Forschungsstiftung, Zweigniederlassung Protein tyrosine phosphatase, non-receptor type 11 (ptpn11) and triple-negative breast cancer
US9244074B2 (en) 2011-06-07 2016-01-26 University Of Hawaii Biomarker of asbestos exposure and mesothelioma
WO2012170742A2 (en) 2011-06-07 2012-12-13 University Of Hawaii Treatment and prevention of cancer with hmgb1 antagonists
EP2721160A2 (en) 2011-06-14 2014-04-23 Synthon Biopharmaceuticals B.V. Compositions and methods for making and biocontaining auxotrophic transgenic plants
WO2012172495A1 (en) 2011-06-14 2012-12-20 Novartis Ag Compositions and methods for antibodies targeting tem8
MX2013015174A (es) 2011-06-21 2014-09-22 Pioneer Hi Bred Int Metodos y composiciones para producir plantas esteriles masculinas.
CN103620029B (zh) 2011-06-24 2017-06-09 诺维信公司 具有蛋白酶活性的多肽和编码它们的多核苷酸
DK2726651T3 (en) 2011-06-28 2019-01-28 Codexis Inc PROTEIN INVARIANT GENERATION BY REGION SHUFFLING
EP2540824A1 (en) 2011-06-30 2013-01-02 The Procter & Gamble Company Cleaning compositions comprising amylase variants reference to a sequence listing
EP3421595B1 (en) 2011-06-30 2020-10-07 Novozymes A/S Alpha-amylase-variants
CN103703124B (zh) 2011-06-30 2021-01-15 诺维信公司 用于筛选α-淀粉酶的方法
US8778652B2 (en) 2011-06-30 2014-07-15 Codexis, Inc. Pentose fermentation by a recombinant microorganism
WO2013006756A2 (en) 2011-07-06 2013-01-10 Novozymes A/S Alpha amylase variants and polynucleotides encoding same
CN103703136A (zh) 2011-07-07 2014-04-02 诺维信公司 采用真菌过氧化酶的脂族烃的区域选择性氧化
EP2729159B1 (en) 2011-07-08 2017-05-24 Defensin Therapeutics ApS Oral treatment of inflammatory bowel disease
WO2013009935A2 (en) 2011-07-12 2013-01-17 Two Blades Foundation Late blight resistance genes
CN103827298A (zh) 2011-07-15 2014-05-28 诺维信公司 脂肪酶变体及其编码多核苷酸
WO2013012643A1 (en) 2011-07-15 2013-01-24 Syngenta Participations Ag Polynucleotides encoding trehalose-6-phosphate phosphatase and methods of use thereof
WO2013012788A2 (en) 2011-07-15 2013-01-24 Syngenta Participations Ag Methods of increasing yield and stress tolerance in a plant
CN103703139A (zh) 2011-07-22 2014-04-02 诺维信北美公司 用于预处理纤维素材料和改进其水解的方法
US9322007B2 (en) 2011-07-22 2016-04-26 The California Institute Of Technology Stable fungal Cel6 enzyme variants
WO2013015993A1 (en) 2011-07-28 2013-01-31 Syngenta Participations Ag Methods and compositions for controlling nematode pests
UY34227A (es) 2011-07-28 2013-02-28 Athenix Corp Acido nucleico recombinante que codifica el gen de la toxina axmi270, vectores, células, plantas y sus métodos de empleo
EA033285B1 (ru) 2011-07-28 2019-09-30 Атеникс Корп. Варианты белков axmi205 и способы их применения
EP2737068A1 (en) 2011-07-29 2014-06-04 Athenix Corp. Axmi279 pesticidal gene and methods for its use
CA2844101A1 (en) 2011-08-03 2013-02-07 E. I. Du Pont De Nemours And Company Methods and compositions for targeted integration in a plant
EP2739728B1 (en) 2011-08-04 2017-07-12 Novozymes A/S Polypeptides having endoglucanase activity and polynucleotides encoding same
EP2739727B1 (en) 2011-08-04 2016-05-25 Novozymes, Inc. Polypeptides having xylanase activity and polynucleotides encoding same
DK2739311T3 (en) 2011-08-04 2018-04-23 Amgen Inc Method of treating bone slit defects
WO2013021065A1 (en) 2011-08-10 2013-02-14 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
WO2013021062A1 (en) 2011-08-10 2013-02-14 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
WO2013021059A1 (en) 2011-08-10 2013-02-14 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
US9382559B2 (en) 2011-08-10 2016-07-05 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
CN103732740B (zh) 2011-08-10 2018-07-31 诺维信公司 具有过氧化酶活性的多肽及编码该多肽的多核苷酸
US9506044B2 (en) 2011-08-10 2016-11-29 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
WO2013021064A1 (en) 2011-08-10 2013-02-14 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
US9000138B2 (en) 2011-08-15 2015-04-07 Novozymes A/S Expression constructs comprising a Terebella lapidaria nucleic acid encoding a cellulase, host cells, and methods of making the cellulase
WO2013024156A2 (en) 2011-08-17 2013-02-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Combinations of anti-hcv-entry factor antibodies and interferons for the treatment and the prevention of hcv infection
WO2013024157A2 (en) 2011-08-17 2013-02-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Combinations of host targeting agents for the treatment and the prevention of hcv infection
WO2013024155A1 (en) 2011-08-17 2013-02-21 Inserm (Institut National De La Sante Et De La Recherche Medicale) Combinations of anti-hcv-entry factor antibodies and direct acting antivirals for the treatment and the prevention of hcv infection
US20130052213A1 (en) 2011-08-19 2013-02-28 Novozymes A/S Novel immunomodulatory peptide
BR112014003444A2 (pt) 2011-08-19 2017-03-14 Novozymes Inc célula hospedeira recombinante, e, método para produzir um ácido c4-dicarboxílico
EP2744900B1 (en) 2011-08-19 2017-07-19 Novozymes A/S Polypeptides having protease activity
DK2748317T3 (en) 2011-08-22 2017-07-17 Codexis Inc GH61 glycoside hydrolase protein variants and cofactors that enhance GH61 activity
WO2013029496A1 (en) 2011-08-26 2013-03-07 Novozymes A/S Polypeptides having glucoamylase activity and polynucleotides encoding same
US11377663B1 (en) 2011-08-30 2022-07-05 Monsanto Technology Llc Genetic regulatory elements
HUE037829T2 (hu) 2011-08-31 2018-09-28 St Jude Childrens Res Hospital Eljárások és készítmények lizoszomális exocitózis aktivitás mértékének detektálására és alkamazási eljárások
DK2748315T3 (en) 2011-09-06 2018-02-05 Novozymes North America Inc GLUCOAMYLASE VARIETIES AND POLYNUCLEOTIDES CODING THEM
CN103998461B (zh) 2011-09-08 2018-02-02 科德克希思公司 用于合成取代的内酰胺的生物催化剂和方法
US9994834B2 (en) 2011-09-09 2018-06-12 Novozymes A/S Polynucleotides encoding polypeptides having alpha-amylase activity and methods of making the same
CA2847879C (en) 2011-09-09 2020-06-23 Novozymes A/S Improving properties of paper materials
WO2013039776A1 (en) 2011-09-13 2013-03-21 Novozymes North America, Inc. Methods of hydrolyzing and fermenting cellulosic material
WO2013043910A1 (en) 2011-09-20 2013-03-28 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
ES2628190T3 (es) 2011-09-22 2017-08-02 Novozymes A/S Polipéptidos con actividad de proteasa y polinucleótidos que codifican los mismos
MX363042B (es) 2011-09-23 2019-03-06 Novozymes As Modificacion del color de textiles.
CA2850070A1 (en) 2011-09-30 2013-04-04 Novozymes A/S Polypeptides having alpha-amylase activity and polynucleotides encoding same
BR112014007719A2 (pt) 2011-09-30 2017-06-13 Codexis Inc proteases fúngicas
BR112014007647A2 (pt) 2011-09-30 2017-04-18 Novozymes As “variante de 3-hpdh, polipeptídeo tendo atividade de 3-hpdh, polinucleotídeo isolado, célula hospedeira microbiana transgênica, e, método para produzir 3-hp
EP2764093B1 (en) 2011-10-05 2018-04-18 The Rockefeller University Dimeric bacteriophage lysins
US9353362B2 (en) 2011-10-11 2016-05-31 Novozymes A/S Glucoamylase variants and polynucleotides encoding same
RU2019133467A (ru) 2011-10-11 2020-07-06 МЕДИММЬЮН, ЭлЭлСи Cd40l-специфичные каркасные структуры, происходящие из tn3, и способы их применения
CN103890170B (zh) 2011-10-17 2018-02-02 诺维信公司 α‑淀粉酶变体以及对其进行编码的多核苷酸
EP2768957B1 (en) 2011-10-17 2018-05-09 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
BR112014009954A2 (pt) 2011-10-28 2017-12-05 Du Pont constructo, célula, planta, semente e método
DK2773656T3 (da) 2011-10-31 2019-09-09 Novozymes Inc Polypeptider med cellulolyseforbedrende aktivitet og polynukleotider, som koder for dem
US9221906B2 (en) 2011-11-01 2015-12-29 Bionomics Inc. Methods of inhibiting solid tumor growth by administering GPR49 antibodies
WO2013067060A1 (en) 2011-11-01 2013-05-10 Bionomics, Inc. Anti-gpr49 antibodies
AU2012332588B2 (en) 2011-11-01 2017-09-07 Bionomics, Inc. Methods of blocking cancer stem cell growth
WO2013067057A1 (en) 2011-11-01 2013-05-10 Bionomics, Inc. Anti-gpr49 antibodies
WO2013064195A1 (en) 2011-11-04 2013-05-10 Enel Ingegneria E Ricerca S.P.A. A new heat-stable carbonic anhydrase and uses thereof
KR102052774B1 (ko) 2011-11-04 2019-12-04 자임워크스 인코포레이티드 Ｆｃ 도메인 내의 돌연변이를 갖는 안정한 이종이합체 항체 설계
EP2776838A1 (en) 2011-11-08 2014-09-17 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research Early diagnostic of neurodegenerative diseases
US20140314787A1 (en) 2011-11-08 2014-10-23 Novartis Forschungsstiftung, Zweigniederlassung, Friedrich Miescher Institute Treatment for neurodegenerative diseases
EP2794873A1 (en) 2011-11-08 2014-10-29 Novozymes A/S Methods for production of archeae protease in yeast
CN104053771B (zh) 2011-11-18 2016-11-09 科德克希思公司 用于制备羟基取代的氨基甲酸酯的生物催化剂
BR112014011902A2 (pt) 2011-11-18 2017-05-16 Novozymes As polipeptídeo e polinucleotídeo isolados, célula hospedeira recombinante, métodos para produzir um polipeptídeo, e para produzir um mutante de uma célula parental, processos para produzir uma proteína, para degradar ou converter um material celulósico ou material contendo xilano, para produzir um produto de fermentação, e para fermentar um material celulósico ou material contendo xilano, e, formulação de caldo integral ou composição de cultura de células
WO2013075066A2 (en) 2011-11-18 2013-05-23 Eleven Biotherapeutics, Inc. Proteins with improved half-life and other properties
WO2013119302A2 (en) 2011-11-21 2013-08-15 Novozymes, Inc. Gh61 polypeptide variants and polynucleotides encoding same
BR112014011387A2 (pt) 2011-11-22 2017-05-02 Novozymes Inc polipeptídeo e polinucleotídeo isolados, célula hospedeira recombinante, métodos para produzir um polipeptídeo, um mutante de uma célula precursora, e uma proteína, e para inibir a expressão de um polipeptídeo, planta transgênica, parte da planta ou célula de planta, molécula de rna, processos para degradar ou converter um material celulósico ou contendo xilano, para sintetizar um produto de fermentação, e para fermentar um material celulósico ou contendo xilano, e, formulação de caldo completo ou composição de cultura de célula
CN103958657A (zh) 2011-11-25 2014-07-30 诺维信公司 枯草杆菌酶变体以及编码该枯草杆菌酶变体的多核苷酸
MX353710B (es) 2011-11-25 2018-01-25 Novozymes As Polipeptidos que tienen actividad de lisozima y polinucleotidos que los codifican.
WO2013079015A1 (en) 2011-12-01 2013-06-06 Novozymes, Inc. Polypeptides having beta-xylosidase activity and polynucleotides encoding same
WO2013079533A1 (en) 2011-12-02 2013-06-06 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
US9169469B2 (en) 2011-12-02 2015-10-27 Novozymes A/S Polypeptides having peroxygenase activity and polynucleotides encoding same
EA201491107A1 (ru) 2011-12-05 2014-11-28 Новартис Аг Антитела к рецептору эпидермального фактора роста 3 (her3), направленные на домен ii her3
CN104159924B (zh) 2011-12-05 2018-03-16 诺华股份有限公司 表皮生长因子受体3(her3)的抗体
EP3272862A1 (en) 2011-12-16 2018-01-24 Novozymes, Inc. Polypeptides having laccase activity and polynucleotides encoding same
EP2794870A4 (en) 2011-12-19 2015-06-17 Novozymes Inc POLYPEPTIDES WITH XYLANASE ACTIVITY AND POLYNUCLEOTIDES THAT CODE
WO2013096244A1 (en) 2011-12-20 2013-06-27 Codexis, Inc. Endoglucanase 1b (eg1b) variants
EP2607468A1 (en) 2011-12-20 2013-06-26 Henkel AG & Co. KGaA Detergent compositions comprising subtilase variants
JP2015504660A (ja) 2011-12-20 2015-02-16 ノボザイムス アクティーゼルスカブ サブチラーゼ変異体およびそれをコードするポリヌクレオチド
US10036050B2 (en) 2011-12-20 2018-07-31 Novozymes, Inc. Cellobiohydrolase variants and polynucleotides encoding same
EP2794656B1 (en) 2011-12-21 2019-02-27 Novartis AG Compositions comprising antibodies targeting factor p and c5
BR112014015564B8 (pt) 2011-12-22 2022-11-08 Du Pont Construtos de dna recombinante e métodos de aumento do teor de óleo de sementes de soja
ES2721882T3 (es) 2011-12-23 2019-08-06 Pfizer Regiones constantes de anticuerpo modificadas por ingeniería genética para conjugación específica de sitio y procedimientos y usos de las mismas
CN107126560A (zh) 2011-12-28 2017-09-05 安进公司 通过使用抗骨硬化蛋白抗体治疗牙槽骨流失的方法
US10174301B2 (en) 2011-12-28 2019-01-08 Novozymes A/S Methods for improving the nutritional value of the animal feed using a protease
WO2013102825A1 (en) 2012-01-02 2013-07-11 Novartis Ag Cdcp1 and breast cancer
CN104039819A (zh) 2012-01-06 2014-09-10 先锋国际良种公司 用于植物繁殖组织中序列表达的组合物和方法
CA2860692A1 (en) 2012-01-06 2013-07-11 Pioneer Hi-Bred International, Inc. A method to screen plants for genetic elements inducing parthenogenesis in plants
WO2013103365A1 (en) 2012-01-06 2013-07-11 Pioneer Hi-Bred International, Inc. Pollen preferred promoters and methods of use
EP2800816A1 (en) 2012-01-06 2014-11-12 Pioneer Hi-Bred International Inc. Ovule specific promoter and methods of use
WO2013104660A1 (en) 2012-01-13 2013-07-18 Dupont Nutrition Biosciences Aps Process for treating a plant oil comprising hydrolysing chlorophyll or a chlorophyll derivative and involving partial caustic neutralisation
WO2013104659A2 (en) 2012-01-13 2013-07-18 Dupont Nutrition Biosciences Aps Process
MX2014008764A (es) 2012-01-26 2014-08-27 Novozymes As Uso de polipeptidos que tienen actividad proteasa en alimentos para animales y detergentes.
WO2013111018A1 (en) 2012-01-26 2013-08-01 Norfolk Plant Sciences, Ltd. Methods for increasing the anthocyanin content of citrus fruit
AR089793A1 (es) 2012-01-27 2014-09-17 Du Pont Metodos y composiciones para generar locus de rasgos transgenicos complejos
AR090418A1 (es) 2012-02-01 2014-11-12 Dow Agrosciences Llc Peptido de transito al cloroplasto
BR122021018583B1 (pt) 2012-02-03 2022-09-06 The Procter & Gamble Company Método para limpar um material têxtil ou uma superfície dura ou outra superfície em cuidados com tecidos e com a casa
ES2794644T3 (es) 2012-02-03 2020-11-18 Novozymes As Variantes de lipasa y polinucleótidos que las codifican
EP2628785B1 (en) 2012-02-17 2016-05-18 Henkel AG & Co. KGaA Detergent compositions comprising subtilase variants
WO2013120948A1 (en) 2012-02-17 2013-08-22 Novozymes A/S Subtilisin variants and polynucleotides encoding same
PT2817325T (pt) 2012-02-20 2019-10-01 Novozymes As Polipéptidos que têm atividade da endoglucanase e polinucleotídeos codificando mesmos
GB201203374D0 (en) 2012-02-28 2012-04-11 Univ Sheffield Alkane production
BR112014022034A2 (pt) 2012-03-08 2017-07-11 Athenix Corp gene da toxina axmi335 de bacillus thuringiensis e métodos para a sua utilização
AR090275A1 (es) 2012-03-08 2014-10-29 Athenix Corp Gen de la delta-endotoxina axmi345 y sus metodos de empleo
IN2014DN06908A (zh) 2012-03-14 2015-05-15 Du Pont
CN104168760B (zh) 2012-03-14 2018-06-22 纳幕尔杜邦公司 编码fasciated ear3(fea3)的核苷酸序列及其使用方法
CA2867342A1 (en) 2012-03-14 2013-09-19 Iowa State University Research Foundation, Inc. Nucleotide sequences encoding fasciated ear4 (fea4) and methods of use thereof
ES2664328T3 (es) 2012-03-16 2018-04-19 Albumedix A/S Variantes de albúmina
DK2828385T3 (en) 2012-03-23 2018-03-12 Codexis Inc BIOCATALIZATORS AND METHODS FOR SYNTHETIZING DERIVATIVES AND TRYPTAMIN AND TRYPTAMINE ANALOGS
US20150266961A1 (en) 2012-03-29 2015-09-24 Novartis Forschungsstiftung, Zweigniederlassung, Fridrich Miescher Institute Inhibition of interleukin-8 and/or its receptor cxcr1 in the treatment of her2/her3-overexpressing breast cancer
CN104204212A (zh) 2012-03-31 2014-12-10 诺维信公司 使用环化酶进行环氧化
CN107988181A (zh) 2012-04-02 2018-05-04 诺维信公司 脂肪酶变体以及编码其的多核苷酸
US9394556B2 (en) 2012-04-23 2016-07-19 Novozymes A/S Polypeptides having glucuronyl esterase activity and polynucleotides encoding same
BR112014026268A2 (pt) 2012-04-23 2017-07-18 Novozymes As polipeptídeo isolado, composição, polinucleotídeo isolado, construção de ácido nucleico ou vetor de expressão, célula hospedeira recombinante, e, métodos de produção de um polipeptídeo e de degradação ou conversão de um material celulósico
CA2870773A1 (en) 2012-04-25 2013-10-31 Novozymes A/S Method of baking
WO2013160372A1 (en) 2012-04-27 2013-10-31 Dupont Nutrition Biosciences Aps Process for treating plant oil involving addition of serial doses of chlorophyll or chlorophyll derivative degrading enzyme
WO2013160374A1 (en) 2012-04-27 2013-10-31 Dupont Nutrition Biosciences Aps Process for refining crude plant oil involving enzymatic hydrolysis and gum recycling
CA2868308A1 (en) 2012-04-27 2013-10-31 Novozymes, Inc. Gh61 polypeptide variants and polynucleotides encoding same
CA2872096A1 (en) 2012-05-04 2013-11-07 E. I. Du Pont De Nemours And Company Compositions and methods comprising sequences having meganuclease activity
ES2643216T3 (es) 2012-05-07 2017-11-21 Novozymes A/S Polipéptidos con actividad de degradación de xantano y polinucleótidos que codifican la misma
ES2712682T3 (es) 2012-05-08 2019-05-14 Codexis Inc Biocatalizadores y métodos para la hidroxilación de compuestos químicos
JP6351572B2 (ja) 2012-05-10 2018-07-04 ザイムワークス，インコーポレイテッド Ｆｃドメインに突然変異を有する免疫グロブリン重鎖のヘテロ多量体構築物
WO2013170050A1 (en) 2012-05-11 2013-11-14 Codexis, Inc. Engineered imine reductases and methods for the reductive amination of ketone and amine compounds
CN104379734B (zh) 2012-05-18 2018-05-15 诺维信公司 具有改进的转化效率的细菌突变体
WO2013178699A1 (en) 2012-05-31 2013-12-05 Novozymes A/S Isopropanol production by bacterial hosts
CN109234250A (zh) 2012-05-31 2019-01-18 诺维信公司 具有有机磷水解酶活性的多肽
CN104334576B (zh) 2012-06-04 2019-02-19 诺华股份有限公司 位点特异性标记方法及由此产生的分子
WO2013188305A2 (en) 2012-06-11 2013-12-19 Codexis, Inc. Fungal beta-xylosidase variants
US20130337442A1 (en) 2012-06-15 2013-12-19 Pioneer Hi-Bred International, Inc. Genetic loci associated with soybean cyst nematode resistance and methods of use
CA2876426A1 (en) 2012-06-15 2013-12-19 E. I. Du Pont De Nemours And Company Methods and compositions involving als variants with native substrate preference
US20150184208A1 (en) 2012-06-19 2015-07-02 Novozymes A/S Enzymatic reduction of hydroperoxides
BR112014031882A2 (pt) 2012-06-20 2017-08-01 Novozymes As uso de um polipeptídeo isolado, polipeptídeo, composição, polinucleotídeo isolado, construto de ácido nucleico ou vetor de expressão, célula hospedeira de expressão recombinante, métodos para produção de um polipeptídeo, para melhoria do valor nutricional de uma ração animal, e para o tratamento de proteínas, uso de pelo menos um polipeptídeo, aditivo de ração animal, ração animal, e, composição detergente
EP2677035A1 (en) 2012-06-22 2013-12-25 BASF Plant Science Company GmbH Plants having enhanced yield-related traits and a method for making the same
US20150337336A1 (en) 2012-06-22 2015-11-26 Phytogene, Inc. Enzymes and methods for styrene synthesis
WO2014001482A1 (en) 2012-06-29 2014-01-03 Novartis Forschungsstiftung, Zweigniererlassung, Friedrich Miescher Institute For Biomedical Research Treating diseases by modulating a specific isoform of mkl1
CA2840683C (en) 2012-06-29 2020-03-10 Athenix Corp. Axmi277 nematode toxin and methods for its use
EP2870242A1 (en) 2012-07-05 2015-05-13 Novartis Forschungsstiftung, Zweigniederlassung Friedrich Miescher Institute For Biomedical Research New treatment for neurodegenerative diseases
WO2014006100A1 (en) 2012-07-05 2014-01-09 Ucb Pharma S.A. Treatment for bone diseases
CN104471048B (zh) 2012-07-12 2018-11-16 诺维信公司 具有脂肪酶活性的多肽及编码它的多核苷酸
BR112014032865A2 (pt) 2012-07-18 2017-08-01 Novozymes As método para tratamento de têxtil de poliéster, e, composição
US20150152452A1 (en) 2012-07-20 2015-06-04 Novozymes A/S Enzymatic Oxidation of 5-Hydroxymethylfurfural and Derivatives Thereof
NZ630363A (en) 2012-07-25 2018-09-28 Celldex Therapeutics Inc Anti-kit antibodies and uses thereof
EP2880159A1 (en) 2012-07-30 2015-06-10 Allylix, Inc. Sclareol and labdenediol diphosphate synthase polypeptides encoding nucleic acid molecules and uses thereof
US10221442B2 (en) 2012-08-14 2019-03-05 10X Genomics, Inc. Compositions and methods for sample processing
US9701998B2 (en) 2012-12-14 2017-07-11 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10323279B2 (en) 2012-08-14 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
US10273541B2 (en) 2012-08-14 2019-04-30 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10752949B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10584381B2 (en) 2012-08-14 2020-03-10 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9951386B2 (en) 2014-06-26 2018-04-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
CA2881685C (en) 2012-08-14 2023-12-05 10X Genomics, Inc. Microcapsule compositions and methods
CN104870640A (zh) 2012-08-16 2015-08-26 孟加拉朱特研究所 来自菜豆壳球孢菌的纤维素和/或半纤维素降解酶及其用途
KR102043363B1 (ko) 2012-08-16 2019-11-11 방글라데시 주트 리서치 인스티튜트 마크로포미나 파세올리나로부터의 펙틴 분해 효소 및 이의 용도
PT2885405T (pt) 2012-08-16 2019-07-19 Novozymes As Método de tratamento de têxtil com endoglucanase
WO2014027062A1 (en) 2012-08-17 2014-02-20 Novozymes A/S Thermostable asparaginase variants and polynucleotides encoding same
CN104603265A (zh) 2012-08-22 2015-05-06 诺维信公司 包括金属蛋白酶的洗涤剂组合物
EP2888360B1 (en) 2012-08-22 2017-10-25 Novozymes A/S Metalloproteases from alicyclobacillus sp.
MX2015002211A (es) 2012-08-22 2015-05-08 Novozymes As Metaloproteasa de exiguobacterium.
WO2014036238A1 (en) 2012-08-30 2014-03-06 Athenix Corp. Axmi-234 and axmi-235 toxin genes and methods for their use
WO2014033266A1 (en) 2012-08-31 2014-03-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-sr-bi antibodies for the inhibition of hepatitis c virus infection
US9771570B2 (en) 2012-09-05 2017-09-26 Novozymes A/S Polypeptides having protease activity
AU2013202506B2 (en) 2012-09-07 2015-06-18 Celgene Corporation Resistance biomarkers for hdac inhibitors
EP2895610B1 (en) 2012-09-13 2019-11-06 Indiana University Research and Technology Corporation Compositions and systems for conferring disease resistance in plants and methods of use thereof
CN104685052A (zh) 2012-09-19 2015-06-03 诺维信股份有限公司 用于增强纤维素材料的降解或转化的方法
EP2898070A2 (en) 2012-09-20 2015-07-29 Novozymes A/S Screening polynucleotide libraries for variants that encode functional proteins
WO2014052630A1 (en) 2012-09-27 2014-04-03 Novozymes, Inc. Bacterial mutants with improved transformation efficiency
US10005988B2 (en) 2012-10-05 2018-06-26 Novozymes A/S Reducing adhesion of bacteria to a surface or releasing bacteria from a surface to which they adhere using endo-beta-A,4-glucanases
US10035996B2 (en) 2012-10-08 2018-07-31 Novozymes, Inc. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
JP2015533832A (ja) 2012-10-09 2015-11-26 アイジェニカ バイオセラピューティクス インコーポレイテッド 抗Ｃ１６ｏｒｆ５４抗体およびその使用方法
US20150259696A1 (en) 2012-10-11 2015-09-17 Shane E. Abbitt Guard cell promoters and uses thereof
US9458435B2 (en) 2012-10-12 2016-10-04 Novozymes A/S Polypeptides having peroxygenase activity
WO2014056920A2 (en) 2012-10-12 2014-04-17 Novozymes A/S Polypeptides having peroxygenase activity
CN104718286B (zh) 2012-10-12 2018-10-30 诺维信公司 具有过氧合酶活性的多肽
EP2906687B1 (en) 2012-10-12 2018-08-15 Novozymes A/S Polypeptides having peroxygenase activity
EP2906686B1 (en) 2012-10-12 2018-08-29 Novozymes A/S Polypeptides having peroxygenase activity
CN104718287B (zh) 2012-10-12 2018-02-06 诺维信公司 具有过氧合酶活性的多肽
CN104704115B (zh) 2012-10-12 2018-11-02 诺维信公司 具有过氧合酶活性的多肽
US9783820B2 (en) 2012-10-15 2017-10-10 Pioneer Hi-Bred International, Inc. Methods and compositions to enhance activity of Cry endotoxins
EP2909230B1 (en) 2012-10-19 2019-05-22 Danisco US Inc. Stabilization of biomimetic membranes
WO2014066497A2 (en) 2012-10-23 2014-05-01 Montana State University Production of high quality durum wheat having increased amylose content
WO2014066141A2 (en) 2012-10-24 2014-05-01 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
US9663532B2 (en) 2012-10-29 2017-05-30 University Of Rochester Artemisinin derivatives, methods for their preparation and their use as antimalarial agents
WO2014068010A1 (en) 2012-10-31 2014-05-08 Novozymes A/S Isopropanol production by bacterial hosts
EP2914729B1 (en) 2012-11-01 2022-01-26 The University of British Columbia Cytochrome p450 and cytochrome p450 reductase polypeptides, encoding nucleic acid molecules and uses thereof
AU2013343503B2 (en) 2012-11-08 2017-12-14 Albumedix Ltd. Albumin variants
AU2013202507B9 (en) 2012-11-14 2015-08-13 Celgene Corporation Inhibition of drug resistant cancer cells
WO2014076232A2 (en) 2012-11-19 2014-05-22 Novozymes A/S Isopropanol production by recombinant hosts using an hmg-coa intermediate
EP2922953A4 (en) 2012-11-20 2016-11-09 Shell Int Research FERMENTATION OF PENTOSIS BY A RECOMBINANT MICROORGANISM
UY35148A (es) 2012-11-21 2014-05-30 Amgen Inc Immunoglobulinas heterodiméricas
CN105209626B (zh) 2012-11-30 2019-09-03 诺维信股份有限公司 通过重组酵母生产3-羟基丙酸
WO2014084859A1 (en) 2012-11-30 2014-06-05 Novartis Ag Molecules and methods for modulating tmem16a activities
MX368067B (es) 2012-12-05 2019-09-18 Novartis Ag Composiciones y métodos para dirigir anticuerpos a la eritropoyetina (epo).
EP2740840A1 (en) 2012-12-07 2014-06-11 Novozymes A/S Improving drainage of paper pulp
EP3556836A1 (en) 2012-12-07 2019-10-23 Novozymes A/S Preventing adhesion of bacteria
DK2928865T3 (en) 2012-12-07 2018-06-18 Merck Sharp & Dohme BIOCATALYTIC TRANSAMINATION PROCEDURE
US20150307871A1 (en) 2012-12-07 2015-10-29 Novozymes A/S Method for generating site-specific mutations in filamentous fungi
US20150368627A1 (en) 2012-12-11 2015-12-24 Novozymes A/S Polypeptides Having Phospholipase C Activity and Polynucleotides Encoding Same
WO2014090765A1 (en) 2012-12-12 2014-06-19 Bayer Cropscience Ag Use of 1-[2-fluoro-4-methyl-5-(2,2,2-trifluoroethylsulfinyl)phenyl]-5-amino-3-trifluoromethyl)-1 h-1,2,4 tfia zole for controlling nematodes in nematode-resistant crops
US10533221B2 (en) 2012-12-14 2020-01-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
EP3567116A1 (en) 2012-12-14 2019-11-13 10X Genomics, Inc. Methods and systems for processing polynucleotides
EP2931885A1 (en) 2012-12-14 2015-10-21 Novozymes A/S Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
WO2014090940A1 (en) 2012-12-14 2014-06-19 Novozymes A/S Removal of skin-derived body soils
US9765317B2 (en) 2012-12-17 2017-09-19 Novozymes A/S Alpha-amylases and polynucleotides encoding same
UY35195A (es) 2012-12-18 2014-07-31 Novartis Ag Composiciones y metodos para proteinas de accion prolongada
WO2014099798A1 (en) 2012-12-19 2014-06-26 Novozymes A/S Polypeptides having cellulolytic enhancinc activity and polynucleotides encoding same
WO2014099730A1 (en) 2012-12-21 2014-06-26 Codexis, Inc. Engineered biocatalysts and methods for synthesizing chiral amines
ES2655032T3 (es) 2012-12-21 2018-02-16 Novozymes A/S Polipéptidos que poseen actividad proteasa y polinucleótidos que los codifican
BR112015014624A2 (pt) 2012-12-24 2017-10-10 Novozymes As polipeptídeo isolado, composição, polinucleotídeo isolado, construto de ácido nucleico ou vetor de expressão, célula hospedeira recombinante, e, método para tratamento de têxtil
CN103013938B (zh) 2012-12-25 2014-12-17 北京大北农科技集团股份有限公司 除草剂抗性蛋白质、其编码基因及用途
CN103060279B (zh) 2012-12-25 2014-08-13 北京大北农科技集团股份有限公司 除草剂抗性蛋白质、其编码基因及用途
CN104903443A (zh) 2013-01-03 2015-09-09 诺维信公司 α-淀粉酶变体以及对其进行编码的多核苷酸
US9540622B2 (en) 2013-01-18 2017-01-10 Codexis, Inc. Engineered biocatalysts useful for carbapenem synthesis
BR112015018449B1 (pt) 2013-01-31 2022-05-31 Codexis, Inc Método implementado por computador para identificar moléculas biológicas tendo uma atividade desejada, meio de armazenamento lido por computador, e, sistema de computação
WO2014120916A1 (en) 2013-02-01 2014-08-07 Bristol-Myers Squibb Company Pegylated domain antibodies monovalent for cd28 binding and methods of use
WO2014122109A1 (en) 2013-02-05 2014-08-14 Novozymes A/S Enzymatic preparation of indigo dyes and intermediates
WO2014122161A2 (en) 2013-02-06 2014-08-14 Novozymes A/S Polypeptides having protease activity
WO2014124258A2 (en) 2013-02-08 2014-08-14 Irm Llc Specific sites for modifying antibodies to make immunoconjugates
PT2953976T (pt) 2013-02-08 2021-06-23 Novartis Ag Sítios específicos de modificação de anticorpos para preparar imunoconjugados
EP2954065B1 (en) 2013-02-08 2021-07-28 10X Genomics, Inc. Partitioning and processing of analytes and other species
WO2014126986A1 (en) 2013-02-13 2014-08-21 Athenix Corp. Use of axmi184 for the control of rootworm insects
JP6557151B2 (ja) 2013-02-20 2019-08-14 エモリー ユニバーシティー 混合物中の核酸を配列決定する方法およびそれに関する組成物
CN105164263B (zh) 2013-02-28 2022-07-01 科德克希思公司 用于工业生物催化的工程化转氨酶多肽
US9487773B2 (en) * 2013-03-01 2016-11-08 Technophage, Investigacao E Desenvolvimento Em Biotecnologia, Sa Cell-based methods for coupling protein interactions and binding molecule selection
CN105247054B (zh) 2013-03-07 2019-06-18 阿则耐克斯公司 毒素基因及其使用方法
CN105164254B (zh) 2013-03-08 2019-06-28 诺维信公司 纤维二糖水解酶变体和编码它们的多核苷酸
BR112015022742A2 (pt) 2013-03-11 2018-11-27 Pionner Hi Bred Int Inc métodos e composições que empregam um domínio de estabilização dependente de sulfonilureia
BR112015022737A2 (pt) 2013-03-11 2018-04-24 Pioneer Hi-Bred International, Inc construto de polinucleotídeo recombinante, célula vegetal, planta, semente, método para regular expressão
US9273322B2 (en) 2013-03-12 2016-03-01 Pioneer Hi Bred International Inc Root-preferred promoter and methods of use
GB201308828D0 (en) 2013-03-12 2013-07-03 Verenium Corp Phytase
US9243258B2 (en) 2013-03-12 2016-01-26 Pioneer Hi Bred International Inc Root-preferred promoter and methods of use
US9803214B2 (en) 2013-03-12 2017-10-31 Pioneer Hi-Bred International, Inc. Breeding pair of wheat plants comprising an MS45 promoter inverted repeat that confers male sterility and a construct that restores fertility
US9498532B2 (en) 2013-03-13 2016-11-22 Novartis Ag Antibody drug conjugates
EP2970479B1 (en) 2013-03-14 2019-04-24 Novartis AG Antibodies against notch 3
WO2014153234A1 (en) 2013-03-14 2014-09-25 Pioneer Hi-Bred International, Inc. Compositions having dicamba decarboxylase activity and methods of use
CA2904537A1 (en) 2013-03-14 2014-09-25 Pioneer Hi-Bred International, Inc. Compositions having dicamba decarboxylase activity and methods of use
GB201308843D0 (en) 2013-03-14 2013-07-03 Verenium Corp Phytase formulation
PT2970934T (pt) 2013-03-14 2017-11-21 Inc Evolva Polipèptidos de valenceno sintase, moléculas de ácido nucleico que os codificam e suas utilizações
US10150800B2 (en) 2013-03-15 2018-12-11 Zyngenia, Inc. EGFR-binding modular recognition domains
WO2014145735A2 (en) 2013-03-15 2014-09-18 Orpro Therapeutics, Inc. Product and process for mucus viscosity normalization
AU2014237003B2 (en) * 2013-03-15 2018-11-15 Gen9, Inc. Compositions and methods for multiplex nucleic acids synthesis
WO2014150914A2 (en) 2013-03-15 2014-09-25 Pioneer Hi-Bred International, Inc. Phi-4 polypeptides and methods for their use
EP3514178A1 (en) 2013-03-15 2019-07-24 Novartis AG Antibody drug conjugates
US9670470B2 (en) 2013-03-21 2017-06-06 Novozymes A/S Polypeptides having phospholipase A activity and polynucleotides encoding same
US9631164B2 (en) 2013-03-21 2017-04-25 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
CN105324483B (zh) 2013-04-18 2018-11-30 科德克希思公司 工程化苯丙氨酸解氨酶多肽
WO2014170218A1 (en) 2013-04-18 2014-10-23 Novozymes A/S Polypeptides having protease activity and polynucleotides encoding same
EP3569611A1 (en) 2013-04-23 2019-11-20 Novozymes A/S Liquid automatic dish washing detergent compositions with stabilised subtilisin
DK2992092T3 (en) 2013-04-30 2018-06-25 Novozymes As GLUCOAMYLASE VARIETIES AND POLYNUCLEOTIDES CODING THEM
US9963690B2 (en) 2013-04-30 2018-05-08 Novozymes A/S Glucoamylase variants and polynucleotides encoding same
CN105164244B (zh) 2013-05-03 2019-08-20 诺维信公司 洗涤剂酶的微囊化
WO2014182990A1 (en) 2013-05-10 2014-11-13 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
EP3418381A1 (en) 2013-05-14 2018-12-26 Novozymes A/S Detergent compositions
CN105209613A (zh) 2013-05-17 2015-12-30 诺维信公司 具有α淀粉酶活性的多肽
SG11201509982UA (zh) 2013-06-06 2016-04-28 Igenica Biotherapeutics Inc
CN114634921A (zh) 2013-06-06 2022-06-17 诺维信公司 α-淀粉酶变体以及对其进行编码的多核苷酸
EP2816115A1 (en) 2013-06-17 2014-12-24 BASF Plant Science Company GmbH Plants having one or more enhanced yield-related traits and a method for making the same
AR096601A1 (es) 2013-06-21 2016-01-20 Novartis Ag Anticuerpos del receptor 1 de ldl oxidado similar a lectina y métodos de uso
UY35620A (es) 2013-06-21 2015-01-30 Novartis Ag Anticuerpos del receptor 1 de ldl oxidado similar a lectina y métodos de uso
WO2014202793A1 (en) 2013-06-21 2014-12-24 Novozymes A/S Production of polypeptides without secretion signal in bacillus
WO2014207224A1 (en) 2013-06-27 2014-12-31 Novozymes A/S Subtilase variants and polynucleotides encoding same
FI3013956T3 (fi) 2013-06-27 2023-05-23 Novozymes As Subtilaasivariantteja ja niitä koodittavia polynukleotideja
AU2014286135A1 (en) 2013-07-04 2015-12-03 Novozymes A/S Polypeptides with xanthan lyase activity having anti-redeposition effect and polynucleotides encoding same
CN105339492A (zh) 2013-07-09 2016-02-17 诺维信公司 具有脂肪酶活性的多肽和编码它们的多核苷酸
US11459579B2 (en) 2013-07-09 2022-10-04 Board Of Trustees Of Michigan State University Transgenic plants produced with a K-domain, and methods and expression cassettes related thereto
BR112015032956B1 (pt) 2013-07-09 2022-09-13 Board Of Trustees Of Michigan State University Método para produzir uma planta transgênica e cassete de expressão
DK3022300T3 (en) 2013-07-17 2018-10-22 Novozymes As : Pullulan chimeras and polynucleotides encoding them
CA2919437A1 (en) 2013-07-25 2015-01-29 Basf Enzymes Llc Phytase
EP3027747B1 (en) 2013-07-29 2018-02-07 Novozymes A/S Protease variants and polynucleotides encoding same
EP3339436B1 (en) 2013-07-29 2021-03-31 Henkel AG & Co. KGaA Detergent composition comprising protease variants
CN117165561A (zh) 2013-07-29 2023-12-05 诺维信公司 蛋白酶变体以及对其进行编码的多核苷酸
EP3027733B1 (en) 2013-07-31 2018-09-12 Novozymes A/S 3-hydroxypropionic acid production by recombinant yeasts expressing an insect aspartate 1-decarboxylase
EP3030902B1 (en) 2013-08-07 2019-09-25 Friedrich Miescher Institute for Biomedical Research New screening method for the treatment friedreich's ataxia
EP3030072B1 (en) 2013-08-08 2020-03-04 Pioneer Hi-Bred International, Inc. Insecticidal polypeptides having broad spectrum activity and uses thereof
UY35696A (es) 2013-08-09 2015-03-27 Athenix Corp ?molécula de adn recombinante que comprende al gen de toxina axmi440, vector, célula huésped, plantas, composiciones y métodos relacionados?.
US9862967B2 (en) 2013-08-09 2018-01-09 Athenix Corp. AXMI281 toxin gene and methods for its use
EP3033420B1 (en) 2013-08-15 2018-02-21 Novozymes A/S Polypeptides having beta-1,3-galactanase activity and polynucleotides encoding same
US10006045B2 (en) 2013-08-16 2018-06-26 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
US10395758B2 (en) 2013-08-30 2019-08-27 10X Genomics, Inc. Sequencing methods
CA2923629A1 (en) 2013-09-11 2015-03-19 Pioneer Hi-Bred International, Inc. Plant regulatory elements and methods of use thereof
CN105705007A (zh) 2013-09-13 2016-06-22 先锋国际良种公司 杀昆虫蛋白及其使用方法
JP6683615B2 (ja) 2013-09-19 2020-04-22 ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー 肝細胞様細胞を産生するための方法および組成物
WO2015048016A2 (en) 2013-09-24 2015-04-02 E. I. Du Pont De Nemours And Company Fasciated inflorescence (fin) sequences and methods of use
CN105765592B (zh) 2013-09-27 2019-12-17 科德克希思公司 用于酶变体的自动筛选的方法、装置和系统
WO2015050959A1 (en) 2013-10-01 2015-04-09 Yale University Anti-kit antibodies and methods of use thereof
WO2015049370A1 (en) 2013-10-03 2015-04-09 Novozymes A/S Detergent composition and use of detergent composition
WO2015059133A1 (en) 2013-10-22 2015-04-30 Novozymes A/S Cellobiose dehydrogenase variants and polynucleotides encoding same
US9822346B2 (en) 2013-11-13 2017-11-21 Codexis, Inc. Engineered imine reductases and methods for the reductive amination of ketone and amine compounds
CN105764343B (zh) 2013-11-25 2020-04-17 拜耳作物科学有限合伙公司 使用axmi-011控制半翅目昆虫
EP2876156A1 (en) 2013-11-26 2015-05-27 Commissariat A L'energie Atomique Et Aux Energies Alternatives New enzymes and method for preparing hydroxylated L-lysine or L-ornithine and analogs thereof
US10683515B2 (en) 2013-11-27 2020-06-16 E. I. Du Pont De Nemours And Company Genetic loci associated with response to abiotic stress
CN105793418A (zh) 2013-11-29 2016-07-20 诺维信公司 过氧合酶变体
US9309314B2 (en) 2013-12-03 2016-04-12 Agency For Science, Technology And Research (A*Star) Polypeptides, nucleic acids and uses thereof
BR122021024287B1 (pt) 2013-12-09 2022-10-11 Athenix Corp Constructo, vetor, célula hospedeira bacteriana, polipeptídeo, composição, método de controle e exterminação de praga lepidóptera, método de proteção de planta e método de produção de polipeptídeo com atividade pesticida
EP3080254A4 (en) 2013-12-11 2017-08-23 Novozymes A/S Cutinase variants and polynucleotides encoding same
US9824068B2 (en) 2013-12-16 2017-11-21 10X Genomics, Inc. Methods and apparatus for sorting data
EP2886656A1 (en) 2013-12-18 2015-06-24 Commissariat A L'energie Atomique Et Aux Energies Alternatives New enzyme and method for preparing 4-hydroxyl benzyl alcohol and derivatives thereof
WO2015094527A1 (en) 2013-12-19 2015-06-25 Danisco Us Inc. Use of hydrophobins to increase gas transferin aerobic fermentation processes
US10030239B2 (en) 2013-12-20 2018-07-24 Novozymes A/S Polypeptides having protease activity and polynucleotides encoding same
EP3087088B1 (en) 2013-12-23 2022-05-25 University Of Rochester Methods and compositions for ribosomal synthesis of macrocyclic peptides
NZ630311A (en) 2013-12-27 2016-03-31 Celgene Corp Romidepsin formulations and uses thereof
EP2896698A1 (en) 2014-01-17 2015-07-22 BASF Plant Science Company GmbH Plants having one or more enhanced yield-related traits and a method for making the same
EP3415624B1 (en) 2014-01-22 2021-08-04 Novozymes A/S Pullulanase variants and polynucleotides encoding same
EP3097112B1 (en) 2014-01-22 2020-05-13 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
RS60593B1 (sr) 2014-01-24 2020-08-31 Ngm Biopharmaceuticals Inc Antitela koja vezuju domen 2 beta-kloto i postupci za njihovu upotrebu
US10358641B2 (en) 2014-01-28 2019-07-23 Dice Molecules Sv, Llc Monoliths with attached recognition compounds, arrays thereof and uses thereof
WO2015116680A1 (en) 2014-01-30 2015-08-06 Two Blades Foundation Plants with enhanced resistance to phytophthora
RU2021113662A (ru) 2014-02-07 2021-05-31 Пайонир Хай-Бред Интернэшнл, Инк. Инсектицидные белки и способы их применения
EP3102684B1 (en) 2014-02-07 2020-05-06 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
EP2910640B1 (en) 2014-02-25 2018-12-05 Biopract GmbH A method for improving substrate degradation in agricultural biogas plants
GB201403775D0 (en) 2014-03-04 2014-04-16 Kymab Ltd Antibodies, uses & methods
US10155935B2 (en) 2014-03-12 2018-12-18 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
KR20160125515A (ko) 2014-03-12 2016-10-31 노파르티스 아게 면역접합체의 제조를 위한 항체의 변형에 사용되는 특정 부위
WO2015143144A1 (en) 2014-03-19 2015-09-24 Novozymes A/S Method for enhancing activity of an x143 polypeptide
WO2015140275A1 (en) 2014-03-19 2015-09-24 Novozymes A/S Polypeptides having phospholipase c activity and polynucleotides encoding same
US20170015950A1 (en) 2014-04-01 2017-01-19 Novozymes A/S Polypeptides having alpha amylase activity
EP3126509A1 (en) 2014-04-01 2017-02-08 DuPont Nutrition Biosciences ApS Method for increasing crude palm oil yields
CN106536556B (zh) 2014-04-04 2020-02-07 生态学有限公司 结合lgr5的人源化抗体
AU2015243445B2 (en) 2014-04-10 2020-05-28 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
EP3129478B1 (en) 2014-04-10 2019-03-27 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
RU2737535C2 (ru) 2014-04-11 2020-12-01 Новозимс А/С Моющая композиция
US10030215B2 (en) 2014-04-15 2018-07-24 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
WO2015161019A1 (en) 2014-04-16 2015-10-22 Codexis, Inc. Engineered tyrosine ammonia lyase
CA2945877C (en) 2014-04-17 2023-09-05 Boehringer Ingelheim Rcv Gmbh & Co Kg Recombinant host cell for expressing proteins of interest
CA2948369C (en) 2014-04-17 2023-09-26 Boehringer Ingelheim Rcv Gmbh & Co Kg Recombinant host cell engineered to overexpress helper proteins
AR100159A1 (es) 2014-04-22 2016-09-14 Du Pont Genes de anhidrasa carbónica plastídica para el incremento de aceite en semillas con expresión incrementada de dgat
WO2015171603A1 (en) 2014-05-06 2015-11-12 Two Blades Foundation Methods for producing plants with enhanced resistance to oomycete pathogens
US10130703B2 (en) 2014-05-09 2018-11-20 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Vaccines against genital herpes simplex infections
EP3143135B1 (en) 2014-05-15 2019-04-10 Novozymes A/S Compositions comprising polypeptides having phospholipase c activity and use thereof
EP3149178B1 (en) 2014-05-27 2020-07-15 Novozymes A/S Lipase variants and polynucleotides encoding same
CA2950273C (en) 2014-05-30 2022-06-21 Novozymes A/S Variants of gh family 11 xylanase and polynucleotides encoding same
EP2952584A1 (en) 2014-06-04 2015-12-09 Boehringer Ingelheim RCV GmbH & Co KG Improved protein production
EP3155097A1 (en) 2014-06-12 2017-04-19 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
EP3154579A1 (en) 2014-06-13 2017-04-19 Friedrich Miescher Institute for Biomedical Research New treatment against influenza virus
ES2694296T3 (es) 2014-06-17 2018-12-19 Centre National De La Recherche Scientifique (Cnrs) Anticuerpos monoclonales anti-pVHL y usos de los mismos
CN114057857A (zh) 2014-06-20 2022-02-18 豪夫迈·罗氏有限公司 基于chagasin的支架组合物、方法和应用
ES2712402T3 (es) 2014-06-25 2019-05-13 Novozymes As Variantes de xilanasa y polinucleótidos que las codifican
WO2015198240A2 (en) 2014-06-25 2015-12-30 Novartis Ag Compositions and methods for long acting proteins
US20170290876A1 (en) 2014-06-25 2017-10-12 Novartis Ag Compositions and methods for long acting proteins
CN106795553B (zh) 2014-06-26 2021-06-04 10X基因组学有限公司 分析来自单个细胞或细胞群体的核酸的方法
CN106575322B (zh) 2014-06-26 2019-06-18 10X基因组学有限公司 核酸序列装配的方法和系统
US20170121646A1 (en) 2014-07-03 2017-05-04 Novozymes A/S Improved Stabilization of Non-Protease Enzyme
WO2016001450A2 (en) 2014-07-04 2016-01-07 Novozymes A/S Subtilase variants and polynucleotides encoding same
EP3878960A1 (en) 2014-07-04 2021-09-15 Novozymes A/S Subtilase variants and polynucleotides encoding same
EP3167052B1 (en) 2014-07-09 2020-01-01 Codexis, Inc. P450-bm3 variants with improved activity
US10786578B2 (en) 2014-08-05 2020-09-29 Novartis Ag CKIT antibody drug conjugates
PE20170287A1 (es) 2014-08-07 2017-04-05 Novartis Ag Anticuerpos anti-proteina similar a angiopoyetina 4 y metodos de uso
EP3194437B1 (en) 2014-08-07 2021-01-20 Novartis AG Angiopoietin-like 4 (angptl4) antibodies and methods of use
CA2955828A1 (en) 2014-08-08 2016-02-11 Pioneer Hi-Bred International, Inc. Ubiquitin promoters and introns and methods of use
PE20170903A1 (es) 2014-08-12 2017-07-12 Novartis Ag Conjugados de farmacos con anticuerpos anti-cdh6
WO2016028999A1 (en) 2014-08-20 2016-02-25 Novozymes A/S Xyloglucan endotransglycosylase variants and polynucleotides encoding same
CN107075511B9 (zh) 2014-08-27 2021-10-26 新英格兰生物实验室公司 合成子的形成
US9963687B2 (en) 2014-08-27 2018-05-08 New England Biolabs, Inc. Fusion polymerase and method for using the same
CN107002056A (zh) 2014-09-05 2017-08-01 诺维信公司 碳水化合物结合模块变体以及编码它们的多核苷酸
US9732373B2 (en) 2014-09-11 2017-08-15 Promega Corporation Luciferase sequences utilizing infrared-emitting substrates to produce enhanced luminescence
RU2017112324A (ru) 2014-09-12 2018-10-15 Пайонир Хай-Бред Интернэшнл, Инк. Создание сайтов сайт-специфической интеграции для сложных локусов признаков в кукурузе и сое, а также способы применения
EP3197557A1 (en) 2014-09-24 2017-08-02 Friedrich Miescher Institute for Biomedical Research Lats and breast cancer
MA41044A (fr) 2014-10-08 2017-08-15 Novartis Ag Compositions et procédés d'utilisation pour une réponse immunitaire accrue et traitement contre le cancer
WO2016061197A1 (en) 2014-10-16 2016-04-21 Pioneer Hi-Bred International, Inc. Insecticidal polypeptides having improved activity spectrum and uses thereof
CN107108705B (zh) 2014-10-16 2021-05-25 先锋国际良种公司 杀昆虫蛋白及其使用方法
US10480008B2 (en) 2014-10-16 2019-11-19 Poineer Hi-Bred International, Inc. Insecticidal polypeptides having broad spectrum activity and uses thereof
GB201418713D0 (en) 2014-10-21 2014-12-03 Kymab Ltd Bindings Proteins
DK3209788T3 (da) 2014-10-23 2019-08-26 Novozymes As Glucoamylasevarianter og polynukleotider, som koder for dem
CA2965438A1 (en) 2014-10-24 2016-04-28 Dupont Nutrition Biosciences Aps Use of proline tolerant tripeptidyl peptidases in feed additive compositions
US20170306360A1 (en) 2014-10-24 2017-10-26 Danisco Us Inc. Method for producing alcohol by use of a tripeptidyl peptidase
KR20170073667A (ko) 2014-10-29 2017-06-28 10엑스 제노믹스, 인크. 표적화 핵산 서열 분석을 위한 방법 및 조성물
US9975122B2 (en) 2014-11-05 2018-05-22 10X Genomics, Inc. Instrument systems for integrated sample processing
WO2016073610A1 (en) 2014-11-07 2016-05-12 Novozymes A/S Xylanase based bleach boosting
CA2967188A1 (en) 2014-11-14 2016-05-19 Novartis Ag Antibody drug conjugates
CN107075489A (zh) 2014-11-20 2017-08-18 诺维信公司 脂环酸芽孢杆菌变体以及编码它们的多核苷酸
WO2016085916A1 (en) 2014-11-25 2016-06-02 Codexis, Inc. Engineered imine reductases and methods for the reductive amination of ketone and amine compounds
WO2016087327A1 (en) 2014-12-01 2016-06-09 Novozymes A/S Polypeptides having pullulanase activity comprising the x25, x45 and cbm41 domains
EP3227438B1 (en) 2014-12-02 2024-03-27 Novozymes A/S Laccase variants and polynucleotides encoding same
CN107075493B (zh) 2014-12-04 2020-09-01 诺维信公司 枯草杆菌酶变体以及编码它们的多核苷酸
WO2016087619A1 (en) 2014-12-04 2016-06-09 Novozymes A/S Liquid cleaning compositions comprising protease variants
WO2016087401A1 (en) 2014-12-05 2016-06-09 Novozymes A/S Lipase variants and polynucleotides encoding same
PT3333191T (pt) 2014-12-11 2020-12-15 Pf Medicament Anticorpos anti-c10orf54 e suas utilizações
MA41142A (fr) 2014-12-12 2017-10-17 Amgen Inc Anticorps anti-sclérostine et utilisation de ceux-ci pour traiter des affections osseuses en tant qu'élements du protocole de traitement
UA124050C2 (uk) 2014-12-12 2021-07-14 Сінгента Партісіпейшнс Аг Химерний ген, який кодує білок, токсичний для кукурудзяного метелика, та спосіб його застосування
CN107002049A (zh) 2014-12-16 2017-08-01 诺维信公司 具有n‑乙酰基葡萄糖胺氧化酶活性的多肽
WO2016100309A1 (en) 2014-12-16 2016-06-23 Pioneer Hi-Bred International, Inc. Restoration of male fertility in wheat
CN107109388A (zh) 2014-12-19 2017-08-29 诺维信公司 蛋白酶变体以及对其进行编码的多核苷酸
EP3234123B1 (en) 2014-12-19 2020-06-03 Novozymes A/S Protease variants and polynucleotides encoding same
WO2016100910A1 (en) 2014-12-19 2016-06-23 Novozymes A/S Recombinant host cells for the production of 3-hydroxypropionic acid
UY36449A (es) 2014-12-19 2016-07-29 Novartis Ag Composiciones y métodos para anticuerpos dirigidos a bmp6
EP4273238A3 (en) 2014-12-19 2023-12-27 Novozymes A/S Compositions comprising polypeptides having xylanase activity and polypeptides having arabinofuranosidase activity
PL3237621T3 (pl) 2014-12-22 2023-10-30 Codexis, Inc. Warianty ludzkiej alfa-galaktozydazy
CN112126675B (zh) 2015-01-12 2022-09-09 10X基因组学有限公司 用于制备核酸测序文库的方法和系统以及用其制备的文库
CN107209814B (zh) 2015-01-13 2021-10-15 10X基因组学有限公司 用于使结构变异和相位信息可视化的系统和方法
CN107406853A (zh) 2015-02-04 2017-11-28 先锋国际良种公司 新颖的Bt毒素受体和使用方法
CA2975529A1 (en) 2015-02-09 2016-08-18 10X Genomics, Inc. Systems and methods for determining structural variation and phasing using variant call data
AU2016219488A1 (en) 2015-02-09 2017-09-14 Bioconsortia, Inc. Agriculturally beneficial microbes, microbial compositions, and consortia
EP3256577B1 (en) 2015-02-10 2020-08-26 Codexis, Inc. Ketoreductase polypeptides for the synthesis of chiral compounds
AU2016217852B2 (en) 2015-02-12 2019-01-17 Dsm Ip Assets B.V. A method for improving feed digestibility in bovine animals
EP3262188B1 (en) 2015-02-24 2021-05-05 10X Genomics, Inc. Methods for targeted nucleic acid sequence coverage
US10557127B2 (en) 2015-02-24 2020-02-11 Novozymes A/S Cellobiohydrolase variants and polynucleotides encoding same
WO2016137973A1 (en) 2015-02-24 2016-09-01 10X Genomics Inc Partition processing methods and systems
US10294466B2 (en) 2015-02-25 2019-05-21 Danisco Us Inc. Alpha-glucosidase, compositions and methods
CN107406821B (zh) 2015-02-27 2021-10-15 诺维信公司 用于生产3-羟基丙酸的突变宿主细胞
PL3265123T3 (pl) 2015-03-03 2023-03-13 Kymab Limited Przeciwciała, zastosowania i sposoby
CA2977026A1 (en) 2015-03-11 2016-09-15 E.I. Du Pont De Nemours And Company Insecticidal combinations of pip-72 and methods of use
WO2016142536A1 (en) 2015-03-11 2016-09-15 Genencor International B.V. Enzymatic activity of lytic polysaccharide monooxygenase
CN107428817B (zh) 2015-03-12 2022-07-12 免疫医疗有限责任公司 纯化白蛋白融合蛋白的方法
EP3070103A1 (en) 2015-03-19 2016-09-21 Institut Hospitalier Universitaire De Strasbourg Anti-Claudin 1 monoclonal antibodies for the prevention and treatment of hepatocellular carcinoma
CA2975279A1 (en) 2015-03-19 2016-09-22 Pioneer Hi-Bred International, Inc. Methods and compositions for accelerated trait introgression
WO2016146473A1 (en) 2015-03-19 2016-09-22 Novozymes A/S A brewing method
CN107667177A (zh) 2015-04-07 2018-02-06 诺维信公司 用于选择具有增强活性的酶的方法
WO2016164596A2 (en) 2015-04-07 2016-10-13 Novozymes A/S Methods for selecting enzymes having lipase activity
US20180105772A1 (en) 2015-04-10 2018-04-19 Novozymes A/S Detergent composition
CN107567489A (zh) 2015-04-10 2018-01-09 诺维信公司 衣物洗涤方法，dna酶和洗涤剂组合物的用途
BR112017022383A2 (pt) 2015-04-17 2018-07-17 Agbiome Inc polipeptídeo recombinante, composição, molécula de ácido nucleico recombinante, ácido nucleico recombinante, célula hospedeira, construto de dna, vetor, método para controlar uma população de pragas, método para produzir um polipeptídeo, planta, semente transgênica da planta, método para proteger uma planta, método para aumentar o rendimento em uma planta
MA41951A (fr) 2015-04-22 2018-02-28 Agbiome Inc Gènes insecticides et procédés d'utilisation
WO2016179522A1 (en) 2015-05-06 2016-11-10 Pioneer Hi-Bred International, Inc. Methods and compositions for the production of unreduced, non-recombined gametes and clonal offspring
CN107531762A (zh) 2015-05-07 2018-01-02 科德克希思公司 青霉素g酰化酶
EP3294883A1 (en) 2015-05-08 2018-03-21 Novozymes A/S Alpha-amylase variants having improved performance and stability
AU2016259703B2 (en) 2015-05-08 2021-12-23 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
AR105416A1 (es) 2015-05-08 2017-10-04 Novozymes As VARIANTES DE a-AMILASA Y POLINUCLEÓTIDOS QUE LAS CODIFICAN
CA2985458A1 (en) 2015-05-09 2016-11-17 Two Blades Foundation Late blight resistance gene from solanum americanum and methods of use
CN108064233B (zh) 2015-05-19 2022-07-15 先锋国际良种公司 杀昆虫蛋白及其使用方法
US10526597B2 (en) 2015-05-21 2020-01-07 Full Spectrum Genetics, Inc. Method of improving characteristics of proteins
CN107532154B (zh) 2015-05-27 2022-09-27 国投生物科技投资有限公司 具有纤维二糖水解酶活性的多肽以及对其进行编码的多核苷酸
WO2016196202A1 (en) 2015-05-29 2016-12-08 Novozymes A/S Polypeptides having protease activity and polynucleotides encoding same
CA2987598A1 (en) 2015-06-03 2016-12-08 AgBiome, Inc. Pesticidal genes and methods of use
US10450539B2 (en) 2015-06-04 2019-10-22 Novozymes A/S Use of M4 metalloprotease in wort production
TN2017000417A1 (en) 2015-06-05 2019-01-16 Novartis Ag Antibodies targeting bone morphogenetic protein 9 (bmp9) and methods therefor
EP3310908B1 (en) 2015-06-16 2020-08-05 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
CA2985490A1 (en) 2015-06-17 2016-12-22 E.I. Du Pont De Nemours And Company Plant regulatory elements and methods of use thereof
EP3310813A1 (en) 2015-06-17 2018-04-25 Novartis AG Antibody drug conjugates
EP3872175A1 (en) 2015-06-18 2021-09-01 Novozymes A/S Subtilase variants and polynucleotides encoding same
WO2016205127A1 (en) 2015-06-18 2016-12-22 Novozymes A/S Polypeptides having trehalase activity and the use thereof in process of producing fermentation products
EP3106508B1 (en) 2015-06-18 2019-11-20 Henkel AG & Co. KGaA Detergent composition comprising subtilase variants
US20160366881A1 (en) 2015-06-22 2016-12-22 AgBiome, Inc. Pesticidal Genes and Methods of Use
ES2879249T3 (es) 2015-06-24 2021-11-22 Genencor Int Bv Polipéptidos que tienen actividad de desmetilación
US10851399B2 (en) 2015-06-25 2020-12-01 Native Microbials, Inc. Methods, apparatuses, and systems for microorganism strain analysis of complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
CN107849597B (zh) 2015-06-25 2022-07-26 原生微生物股份有限公司 基于分析异质微生物群落形成活性微生物菌株的群的方法
US9938558B2 (en) 2015-06-25 2018-04-10 Ascus Biosciences, Inc. Methods, apparatuses, and systems for analyzing microorganism strains from complex heterogeneous communities, predicting and identifying functional relationships and interactions thereof, and selecting and synthesizing microbial ensembles based thereon
US10632157B2 (en) 2016-04-15 2020-04-28 Ascus Biosciences, Inc. Microbial compositions and methods of use for improving fowl production
WO2016207373A1 (en) 2015-06-26 2016-12-29 Novozymes A/S Polypeptides having peroxygenase activity
JOP20200312A1 (ar) 2015-06-26 2017-06-16 Novartis Ag الأجسام المضادة للعامل xi وطرق الاستخدام
CN107849548A (zh) 2015-06-26 2018-03-27 诺维信公司 生物整理系统
EP3313193B1 (en) 2015-06-26 2021-03-24 Novozymes A/S Method for producing a coffee extract
BR112017028035A2 (pt) 2015-06-26 2018-08-28 Dupont Nutrition Biosci Aps aminopeptidases para hidrolisados de proteína
EP3929285A3 (en) 2015-07-01 2022-05-25 Novozymes A/S Methods of reducing odor
MX2017016500A (es) 2015-07-02 2018-03-12 Novozymes As Composiciones de pienso para animales y uso de las mismas.
CN114292829A (zh) 2015-07-06 2022-04-08 诺维信公司 脂肪酶变体以及编码它们的多核苷酸
EP3319987B1 (en) 2015-07-07 2021-05-05 Codexis, Inc. Novel p450-bm3 variants with improved activity
US10577594B2 (en) 2015-07-24 2020-03-03 Novozymes, Inc. Polypeptides having arabinofuranosidase activity and polynucleotides encoding same
US20180216089A1 (en) 2015-07-24 2018-08-02 Novozymes, Inc. Polypeptides Having Beta-Xylosidase Activity And Polynucleotides Encoding Same
MX2018000780A (es) 2015-07-25 2018-03-23 Bioconsortia Inc Microbios, composiciones y consorcios microbianos beneficiosos para la agricultura.
CN115043944A (zh) 2015-08-03 2022-09-13 诺华股份有限公司 治疗fgf21相关病症的方法
CN109475096B (zh) 2015-08-06 2022-08-23 先锋国际良种公司 植物来源的杀昆虫蛋白及其使用方法
CN108137674B (zh) 2015-08-20 2022-12-06 阿尔布梅迪克斯医疗有限公司 白蛋白变体和缀合物
EP3341475A1 (en) 2015-08-24 2018-07-04 Novozymes A/S Beta-alanine aminotransferases for the production of 3-hydroxypropionic acid
AU2016321204B2 (en) 2015-09-08 2022-12-01 Cold Spring Harbor Laboratory Genetic copy number determination using high throughput multiplex sequencing of smashed nucleotides
EP3347377B1 (en) 2015-09-09 2021-02-17 Novartis AG Thymic stromal lymphopoietin (tslp)-binding antibodies and methods of using the antibodies
TWI799366B (zh) 2015-09-15 2023-04-21 美商建南德克公司 胱胺酸結骨架平臺
EP3353195B1 (en) 2015-09-22 2021-11-10 Novozymes A/S Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
AU2016326867B2 (en) 2015-09-22 2018-09-13 F. Hoffmann-La Roche Ag OmpG variants
AU2016332725A1 (en) 2015-09-29 2018-03-22 Celgene Corporation PD-1 binding proteins and methods of use thereof
EP3763818A1 (en) 2015-10-06 2021-01-13 Pierce Biotechnology, Inc. Devices and methods for producing proteins
US10954497B2 (en) 2015-10-07 2021-03-23 Novozymes A/S Polypeptides
WO2017062790A1 (en) 2015-10-09 2017-04-13 Two Blades Foundation Cold shock protein receptors and methods of use
BR112018007474A2 (pt) 2015-10-14 2018-10-30 Novozymes A/S ?limpeza de membranas de filtração de água?
EP4324919A2 (en) 2015-10-14 2024-02-21 Novozymes A/S Polypeptide variants
EP3362574B1 (en) 2015-10-14 2021-07-07 Novozymes A/S Glucoamylase variants and polynucleotides encoding same
CN108291215A (zh) 2015-10-14 2018-07-17 诺维信公司 具有蛋白酶活性的多肽以及编码它们的多核苷酸
WO2017070219A1 (en) 2015-10-20 2017-04-27 Novozymes A/S Lytic polysaccharide monooxygenase (lpmo) variants and polynucleotides encoding same
US20180348224A1 (en) 2015-10-28 2018-12-06 Friedrich Miescher Institute For Biomedical Resear Ch Tenascin-w and biliary tract cancers
US11028346B2 (en) 2015-10-28 2021-06-08 Novozymes A/S Detergent composition comprising protease and amylase variants
ITUB20155272A1 (it) 2015-11-02 2017-05-02 Scuola Normale Superiore Intracellular antibody
US11739343B2 (en) 2015-11-03 2023-08-29 The Sainsbury Laboratory Wheat stripe rust resistance genes and methods of use
BR112018007796A2 (pt) 2015-11-06 2018-10-30 Du Pont plantas de soja, partes de plantas de soja ou sementes de soja, método para selecionar uma célula de soja, métodos de seleção de uma célula de soja e de produção de um locus e molécula de ácido nucleico
CN108473974A (zh) 2015-11-24 2018-08-31 诺维信公司 具有蛋白酶活性的多肽以及编码其的多核苷酸
CN108473999B (zh) 2015-11-30 2022-12-23 先锋国际良种公司 植物调节元件及其使用方法
WO2017093318A1 (en) 2015-12-01 2017-06-08 Novozymes A/S Methods for producing lipases
US10774370B2 (en) 2015-12-04 2020-09-15 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
EP3387125B1 (en) 2015-12-07 2022-10-12 Henkel AG & Co. KGaA Dishwashing compositions comprising polypeptides having beta-glucanase activity and uses thereof
US11208649B2 (en) 2015-12-07 2021-12-28 Zymergen Inc. HTP genomic engineering platform
US9988624B2 (en) 2015-12-07 2018-06-05 Zymergen Inc. Microbial strain improvement by a HTP genomic engineering platform
US10035995B2 (en) 2015-12-07 2018-07-31 Eastman Chemical Company CALB variants
KR20180084756A (ko) 2015-12-07 2018-07-25 지머젠 인코포레이티드 코리네박테리움 글루타미컴으로부터의 프로모터
EP3858996B1 (en) 2015-12-07 2022-08-03 Zymergen Inc. Microbial strain improvement by a htp genomic engineering platform
CN109069623A (zh) 2015-12-18 2018-12-21 诺华股份有限公司 靶向CD32b的抗体及其使用方法
EP3390431A1 (en) 2015-12-18 2018-10-24 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
CA3004056C (en) 2015-12-22 2024-01-23 Pioneer Hi-Bred International, Inc. Tissue-preferred promoters and methods of use
UA126850C2 (uk) 2015-12-22 2023-02-15 Аґбайомі, Інк. Поліпептид, що має пестицидну активність проти шкідників lepidopteran
EP3394243A1 (en) 2015-12-22 2018-10-31 Albumedix Ltd. Improved protein expression strains
CN108603185B (zh) 2015-12-30 2022-05-17 诺维信公司 酶变体以及编码它们的多核苷酸
US10293006B2 (en) 2016-01-07 2019-05-21 Ascus Biosciences, Inc. Microbial compositions for improving milk production in ruminants
WO2017125897A1 (en) 2016-01-21 2017-07-27 Novartis Ag Multispecific molecules targeting cll-1
US20210171927A1 (en) 2016-01-29 2021-06-10 Novozymes A/S Beta-glucanase variants and polynucleotides encoding same
SG11201806757XA (en) 2016-02-11 2018-09-27 10X Genomics Inc Systems, methods, and media for de novo assembly of whole genome sequence data
GB201602535D0 (en) 2016-02-12 2016-03-30 Univ Edinburgh Improved flu vaccine yield
US11685911B2 (en) 2016-02-16 2023-06-27 Monaghan Mushroom S Group Fungal polypeptides having lysozyme activity
EP3419992B1 (en) 2016-02-22 2020-11-18 Danisco US Inc. Fungal high-level protein production system
BR112018016889A2 (pt) 2016-02-23 2019-02-12 Da Volterra variantes de beta-lactamase
US10988749B2 (en) 2016-02-23 2021-04-27 Da Volterra Beta-lactamase variants
WO2017147060A1 (en) 2016-02-25 2017-08-31 Dupont Nutrition Biosciences Aps Method for producing a protein hydrolysate employing an aspergillus fumigatus tripeptidyl peptidase
EP3423576B1 (en) 2016-02-29 2021-05-12 Genia Technologies, Inc. Polymerase variants
EP3423577A1 (en) 2016-03-02 2019-01-09 Novozymes A/S Cellobiohydrolase variants and polynucleotides encoding same
GB201604124D0 (en) 2016-03-10 2016-04-27 Ucb Biopharma Sprl Pharmaceutical formulation
US10745487B2 (en) 2016-03-22 2020-08-18 Bionomics Limited Method of treating cancer by administering an anti-LGR5 monoclonal antibody
EP3433358B1 (en) 2016-03-24 2022-07-06 Novozymes A/S Cellobiohydrolase variants and polynucleotides encoding same
US9988641B2 (en) 2016-04-05 2018-06-05 Corn Products Development, Inc. Compositions and methods for producing starch with novel functionality
CN109312333A (zh) 2016-04-07 2019-02-05 诺维信公司 选择具有蛋白酶活性的酶的方法
US11781151B2 (en) 2016-04-14 2023-10-10 Pioneer Hi-Bred International, Inc. Insecticidal CRY1B variants having improved activity spectrum and uses thereof
US10617134B2 (en) 2016-04-15 2020-04-14 Ascus Biosciences, Inc. Methods for improving agricultural production of fowl by administration of microbial compositions or purified strains thereof
WO2017184673A1 (en) 2016-04-19 2017-10-26 Pioneer Hi-Bred International, Inc. Insecticidal combinations of polypeptides having improved activity spectrum and uses thereof
CN109071647B (zh) 2016-04-27 2022-11-22 诺华股份有限公司 抗生长分化因子15的抗体及其用途
US20190192626A1 (en) 2016-04-29 2019-06-27 Defensin Therapeutics Aps Treatment of liver, biliary tract and pancreatic disorders
WO2017186943A1 (en) 2016-04-29 2017-11-02 Novozymes A/S Detergent compositions and uses thereof
CN109415421B (zh) 2016-05-03 2023-02-28 诺维信公司 α-淀粉酶变体以及编码它们的多核苷酸
CA3018384A1 (en) 2016-05-04 2017-11-09 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
JP7128519B2 (ja) 2016-05-05 2022-08-31 コデクシス， インコーポレイテッド ペニシリンｇアシラーゼ
US11186833B2 (en) 2016-05-09 2021-11-30 Novozymes A/S Variant polypeptides with improved performance and use of the same
WO2017197338A1 (en) 2016-05-13 2017-11-16 10X Genomics, Inc. Microfluidic systems and methods of use
EP3246401A1 (en) 2016-05-20 2017-11-22 Commissariat À L'Énergie Atomique Et Aux Énergies Alternatives New fatty acid decarboxylase and its uses
BR112018073875A2 (pt) 2016-05-24 2019-02-26 Novozymes As polipeptídeo isolado, composição, grânulo, aditivo de ração animal, formulação líquida, ração animal, métodos para liberar galactose de material à base de planta, para melhorar um ou mais parâmetros de desempenho de um animal e o valor nutricional de uma ração animal, para preparar uma ração animal e para produzir o polipeptídeo, uso, polinucleotídeo, construto de ácido nucleico ou vetor de expressão, e, célula hospedeira recombinante.
CN109196098A (zh) 2016-05-24 2019-01-11 诺维信公司 包含具有半乳聚糖酶活性的多肽和具有β-半乳糖苷酶活性的多肽的组合物
US10947520B2 (en) 2016-05-24 2021-03-16 Novozymes A/S Compositions comprising polypeptides having galactanase activity and polypeptides having beta-galactosidase activity
US11058129B2 (en) 2016-05-24 2021-07-13 Novozymes A/S Animal feed additives
TW201802121A (zh) 2016-05-25 2018-01-16 諾華公司 抗因子XI/XIa抗體之逆轉結合劑及其用途
WO2017205535A1 (en) 2016-05-27 2017-11-30 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
CN109563450A (zh) 2016-05-31 2019-04-02 诺维信公司 稳定的液体过氧化物组合物
WO2017207762A1 (en) 2016-06-03 2017-12-07 Novozymes A/S Subtilase variants and polynucleotides encoding same
WO2017211803A1 (en) 2016-06-07 2017-12-14 Novozymes A/S Co-expression of heterologous polypeptides to increase yield
EP3469088B1 (en) 2016-06-09 2023-09-06 Codexis, Inc. Biocatalysts and methods for hydroxylation of chemical compounds
US11299723B2 (en) 2016-06-15 2022-04-12 Codexis, Inc. Engineered beta-glucosidases and glucosylation methods
CN110381988A (zh) 2016-06-15 2019-10-25 诺华股份有限公司 使用骨形态发生蛋白6(bmp6)的抑制剂治疗疾病的方法
EP3472186A1 (en) 2016-06-17 2019-04-24 National Hellenic Research Foundation Systems for recombinant protein production
CA3026653A1 (en) 2016-06-24 2017-12-28 Pioneer Hi-Bred International, Inc. Plant regulatory elements and methods of use thereof
BR112018077510A2 (pt) 2016-06-30 2019-04-02 Novozymes As variantes de lipase e composições que compreendem tensoativo e variante de lipase
EP3478845A4 (en) 2016-06-30 2019-07-31 Zymergen, Inc. METHODS OF PRODUCING A GLUCOSE PERMEASE BANK AND USES THEREOF
WO2018005655A2 (en) 2016-06-30 2018-01-04 Zymergen Inc. Methods for generating a bacterial hemoglobin library and uses thereof
WO2018005411A1 (en) 2016-07-01 2018-01-04 Pioneer Hi-Bred International, Inc. Insecticidal proteins from plants and methods for their use
WO2018002261A1 (en) 2016-07-01 2018-01-04 Novozymes A/S Detergent compositions
WO2018002379A2 (en) 2016-07-01 2018-01-04 Novozymes A/S Enzymatic preparation of indigo dyes and in situ dyeing process
JP2019522988A (ja) 2016-07-05 2019-08-22 ノボザイムス アクティーゼルスカブ ペクチン酸リアーゼ変異体およびこれをコードするポリヌクレオチド
WO2018007573A1 (en) 2016-07-08 2018-01-11 Novozymes A/S Detergent compositions with galactanase
CA3029113A1 (en) 2016-07-08 2018-01-11 Novozymes A/S Xylanase variants and polynucleotides encoding same
AU2017294066A1 (en) 2016-07-08 2019-01-17 Novozymes A/S Polypeptides having xylanase activity and polynucleotides encoding same
CN109415708A (zh) 2016-07-13 2019-03-01 宝洁公司 食物芽孢杆菌脱氧核糖核酸酶变体及其用途
WO2018015295A1 (en) 2016-07-18 2018-01-25 Novozymes A/S Lipase variants, polynucleotides encoding same and the use thereof
WO2018017792A1 (en) 2016-07-20 2018-01-25 Novozymes A/S Heat-stable metagenomic carbonic anhydrases and their use
EP3487996B1 (en) 2016-07-21 2024-04-10 Novozymes A/S Serine protease variants and polynucleotides encoding same
MX2019000743A (es) 2016-07-21 2019-05-20 Novozymes As Variantes de serinproteasa y polinucleotidos que las codifican.
WO2018015444A1 (en) 2016-07-22 2018-01-25 Novozymes A/S Crispr-cas9 genome editing with multiple guide rnas in filamentous fungi
WO2018026868A1 (en) 2016-08-01 2018-02-08 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
US10988747B2 (en) 2016-08-24 2021-04-27 Henkel Ag & Co. Kgaa Detergent composition comprising GH9 endoglucanase variants I
CA3032248A1 (en) 2016-08-24 2018-03-01 Novozymes A/S Xanthan lyase variants and polynucleotides encoding same
EP3504331A1 (en) 2016-08-24 2019-07-03 Henkel AG & Co. KGaA Detergent compositions comprising xanthan lyase variants i
EP3504330A1 (en) 2016-08-24 2019-07-03 Novozymes A/S Gh9 endoglucanase variants and polynucleotides encoding same
JP7316662B2 (ja) 2016-08-26 2023-07-28 コデクシス， インコーポレイテッド 操作されたイミンレダクターゼ、ならびにケトン化合物およびアミン化合物の還元的アミノ化のための方法
CA3035896A1 (en) 2016-09-06 2018-03-15 AgBiome, Inc. Pesticidal genes and methods of use
JP2019534858A (ja) 2016-09-09 2019-12-05 ジェネンテック， インコーポレイテッド Ｆｒｉｚｚｌｅｄの選択的ペプチド阻害剤
EP3512941B1 (en) 2016-09-16 2021-03-24 DuPont Nutrition Biosciences ApS Acetolactate decarboxylase variants having improved specific activity
JP2019534859A (ja) 2016-09-19 2019-12-05 セルジーン コーポレイション Ｐｄ−１結合タンパク質を使用して白斑を治療する方法
CN109952317A (zh) 2016-09-19 2019-06-28 细胞基因公司 使用pd-1结合蛋白治疗免疫病症的方法
US10787674B2 (en) 2016-09-20 2020-09-29 22Nd Century Limited, Llc Trichome specific promoters for the manipulation of cannabinoids and other compounds in grandular trichomes
WO2018060475A1 (en) 2016-09-29 2018-04-05 Novozymes A/S Spore containing granule
CA3038972A1 (en) 2016-09-30 2018-04-05 Dow Agrosciences Llc Binary insecticidal cry toxins
WO2018077938A1 (en) 2016-10-25 2018-05-03 Novozymes A/S Detergent compositions
EP3535285B1 (en) 2016-11-01 2022-04-06 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
WO2018085370A1 (en) 2016-11-02 2018-05-11 Novozymes A/S Processes for reducing production of primeverose during enzymatic saccharification of lignocellulosic material
US11779604B2 (en) 2016-11-03 2023-10-10 Kymab Limited Antibodies, combinations comprising antibodies, biomarkers, uses and methods
KR102489902B1 (ko) 2016-11-11 2023-01-19 바이오 래드 래버러토리스 인코오포레이티드 핵산 샘플을 프로세싱하는 방법
EP3541954A1 (en) 2016-11-21 2019-09-25 Novozymes A/S Yeast cell extract assisted construction of dna molecules
CN110072997A (zh) 2016-11-23 2019-07-30 诺维信公司 具有蛋白酶活性的多肽和编码其的多核苷酸
MX2019005835A (es) 2016-11-23 2019-10-30 BASF Agricultural Solutions Seed US LLC Genes de toxinas axmi669 y axmi991 y metodos para su uso.
EP3548615A1 (en) 2016-11-29 2019-10-09 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
CN109963469A (zh) 2016-11-30 2019-07-02 诺维信公司 烘焙的方法
CN110168095A (zh) 2016-12-06 2019-08-23 诺维信公司 用于使用工程化酵母菌株从含有木糖的纤维素基质生产乙醇的改善方法
KR20190108106A (ko) 2016-12-13 2019-09-23 디펜신 테라퓨틱스 에이피에스 폐의 염증 상태를 치료하는 방법
CN110088123B (zh) 2016-12-14 2023-10-20 先锋国际良种公司 杀昆虫蛋白及其使用方法
CA3047121A1 (en) 2016-12-16 2018-06-21 Two Blades Foundation Late blight resistance genes and methods of use
CN110114465A (zh) 2016-12-20 2019-08-09 诺维信公司 用于戊糖发酵的重组酵母菌株
CN110087626B (zh) 2016-12-20 2022-11-22 高露洁-棕榄公司 口腔护理组合物和用于增白牙齿的方法
RU2728608C1 (ru) 2016-12-20 2020-07-30 Колгейт-Палмолив Компани Двухфазные отбеливающие композиции для ухода за полостью рта
EP3541354B1 (en) 2016-12-20 2021-04-28 Colgate-Palmolive Company Oral care compositions and methods for whitening teeth
BR112019012897A2 (pt) 2016-12-22 2020-01-07 BASF Agricultural Solutions Seed US LLC Métodos de controle de população de pragas, de proteção de plantas contra pragas e de aumento da produção em plantas
US10815525B2 (en) 2016-12-22 2020-10-27 10X Genomics, Inc. Methods and systems for processing polynucleotides
MX2019007491A (es) 2016-12-22 2019-09-06 Pioneer Hi Bred Int Proteinas insecticidas y metodos para su uso.
US10011872B1 (en) 2016-12-22 2018-07-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10550429B2 (en) 2016-12-22 2020-02-04 10X Genomics, Inc. Methods and systems for processing polynucleotides
JP7110199B2 (ja) 2016-12-23 2022-08-01 ノバルティス アーゲー 抗第ＸＩ／ＸＩａ因子抗体による処置法
IL308980A (en) 2016-12-23 2024-01-01 Novartis Ag Antibodies against factor XI and methods of their use
JP2020504620A (ja) 2016-12-28 2020-02-13 アスカス バイオサイエンシーズ， インコーポレイテッド 複雑な不均一コミュニティの完全微生物株の解析、その機能的関連性及び相互作用の決定、ならびにそれに基づく生物反応性の改変剤の同定及び合成、のための方法、装置、及びシステム
WO2018129029A1 (en) 2017-01-04 2018-07-12 Immunogen, Inc. Met antibodies and immunoconjugates and uses thereof
US10745681B2 (en) 2017-01-05 2020-08-18 Codexis, Inc. Penicillin-G acylases
UY37570A (es) 2017-01-18 2018-08-31 Bayer Cropscience Lp Uso de bp005 para el control de patógenos de planta
US11279946B2 (en) 2017-01-18 2022-03-22 Basf Argicultural Solutions Seed Us Llc BP005 toxin gene and methods for its use
WO2018140214A1 (en) 2017-01-24 2018-08-02 Pioneer Hi-Bred International, Inc. Nematicidal protein from pseudomonas
WO2018140966A1 (en) 2017-01-30 2018-08-02 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
CN110506117A (zh) 2017-01-30 2019-11-26 农业生物群落股份有限公司 杀虫基因及使用方法
MX2019009093A (es) 2017-02-01 2019-12-05 Procter & Gamble Composiciones de limpieza que comprenden variantes de amilasa.
JP7242535B2 (ja) 2017-02-01 2023-03-20 ノボザイムス アクティーゼルスカブ α－アミラーゼ変異体
CA3050310A1 (en) 2017-02-03 2018-08-09 Codexis, Inc. Engineered glycosyltransferases and steviol glycoside glucosylation methods
JOP20190187A1 (ar) 2017-02-03 2019-08-01 Novartis Ag مترافقات عقار جسم مضاد لـ ccr7
KR102572663B1 (ko) 2017-02-08 2023-09-01 노파르티스 아게 Fgf21 모방 항체 및 이의 용도
WO2018148001A1 (en) 2017-02-08 2018-08-16 Pioneer Hi-Bred International Inc Insecticidal combinations of plant derived insecticidal proteins and methods for their use
KR102573324B1 (ko) 2017-02-13 2023-08-30 코덱시스, 인코포레이티드 조작된 페닐알라닌 암모니아 리아제 폴리펩티드
WO2018150021A1 (en) 2017-02-20 2018-08-23 Novozymes A/S Lipolytic enzyme for use in baking
CA3055530A1 (en) 2017-03-07 2018-09-13 Danisco Us Inc. Thermostable glucoamylase and methods of use, thereof
CN110612352A (zh) 2017-03-29 2019-12-24 贝林格尔·英格海姆Rcv两合公司 具有改变的膜脂组成的重组宿主细胞
US11149233B2 (en) 2017-03-31 2021-10-19 Novozymes A/S Polypeptides having RNase activity
WO2018177936A1 (en) 2017-03-31 2018-10-04 Novozymes A/S Polypeptides having dnase activity
EP3601550A1 (en) 2017-03-31 2020-02-05 Novozymes A/S Polypeptides having dnase activity
WO2018185618A1 (en) 2017-04-03 2018-10-11 Novartis Ag Anti-cdh6 antibody drug conjugates and anti-gitr antibody combinations and methods of treatment
US20200109354A1 (en) 2017-04-04 2020-04-09 Novozymes A/S Polypeptides
WO2018185152A1 (en) 2017-04-04 2018-10-11 Novozymes A/S Polypeptide compositions and uses thereof
WO2018185181A1 (en) 2017-04-04 2018-10-11 Novozymes A/S Glycosyl hydrolases
BR112019021223A2 (pt) 2017-04-11 2020-04-28 Novozymes As variante de glucoamilase, métodos para aumentar a atividade de hidrólise de amido em bruto de uma glucoamilase e para produzir uma variante de glucoamilase, composição, uso de uma variante de glucoamilase, processos para produzir um produto de fermentação e para produzir um produto de xarope, polinucleotídeo codificando a variante de glucoamilase, construto de ácido nucleico, vetor de expressão, e, célula hospedeira.
CA3058757A1 (en) 2017-04-11 2018-10-18 AgBiome, Inc. Pesticidal genes and methods of use
CA3060715A1 (en) 2017-04-24 2018-11-01 Tesaro, Inc. Methods of manufacturing of niraparib
WO2018197520A1 (en) 2017-04-24 2018-11-01 Dupont Nutrition Biosciences Aps Methods and compositions of anti-crispr proteins for use in plants
CA3061133A1 (en) 2017-04-27 2018-11-01 Codexis, Inc. Ketoreductase polypeptides and polynucleotides
UY37708A (es) 2017-04-28 2018-10-31 Ascus Biosciences Inc Métodos para complementar dietas con alto consumo de granos y/o alto valor energético en rumiantes, mediante la administración de un bioensamble sintético de microbios, o cepas purificadas de los mismos
US11078445B2 (en) 2017-05-05 2021-08-03 Novozymes A/S Compositions comprising lipase and sulfite
CA3058095A1 (en) 2017-05-08 2018-11-15 Novozymes A/S Mannanase variants and polynucleotides encoding same
CA3058092A1 (en) 2017-05-08 2018-11-15 Novozymes A/S Mannanase variants and polynucleotides encoding same
WO2018206535A1 (en) 2017-05-08 2018-11-15 Novozymes A/S Carbohydrate-binding domain and polynucleotides encoding the same
DK3635101T3 (da) 2017-05-08 2021-09-20 Codexis Inc Manipulerede ligasevarianter
BR112019023628A2 (pt) 2017-05-11 2020-06-02 Pioneer Hi-Bred International, Inc. Polipeptídeo inseticida recombinante, proteína inseticida quimérica, proteína de fusão, composição agrícola, polinucleotídeo recombinante, construto de dna, planta transgênica, método para inibir o crescimento ou exterminar uma praga de inseto ou população de praga, método para controlar os danos por praga de inseto, método para controlar infestação de praga e método para melhorar o rendimento de uma cultura
JP2020520645A (ja) 2017-05-19 2020-07-16 ザイマージェン インコーポレイテッド Ｎａｄｐｈの増大を導く生合成経路のゲノム工学
EP3625715A4 (en) 2017-05-19 2021-03-17 10X Genomics, Inc. DATA SET ANALYSIS SYSTEMS AND METHODS
WO2018215937A1 (en) 2017-05-24 2018-11-29 Novartis Ag Interleukin-7 antibody cytokine engrafted proteins and methods of use in the treatment of cancer
JOP20190271A1 (ar) 2017-05-24 2019-11-21 Novartis Ag بروتينات مطعّمة بسيتوكين- الجسم المضاد وطرق الاستخدام للاضطرابات المتعلقة بالمناعة
CN110662762A (zh) 2017-05-24 2020-01-07 诺华股份有限公司 抗体细胞因子移植蛋白和用于治疗癌症的方法
WO2018215938A1 (en) 2017-05-24 2018-11-29 Novartis Ag Antibody-cytokine engrafted proteins and methods of use
US10844372B2 (en) 2017-05-26 2020-11-24 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
EP4230746A3 (en) 2017-05-26 2023-11-01 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
BR112019024827A2 (pt) 2017-05-26 2020-06-16 Pioneer Hi-Bred International, Inc. Construto de dna, planta transgênica ou progênie da mesma, composição e método para controlar uma população de pragas de insetos
US11091753B2 (en) 2017-05-31 2021-08-17 Novozymes A/S Xylose fermenting yeast strains and processes thereof for ethanol production
US20200157581A1 (en) 2017-06-02 2020-05-21 Novozymes A/S Improved Yeast For Ethanol Production
WO2018226880A1 (en) 2017-06-06 2018-12-13 Zymergen Inc. A htp genomic engineering platform for improving escherichia coli
JP2020524494A (ja) 2017-06-06 2020-08-20 ザイマージェン インコーポレイテッド ハイスループットトランスポゾン変異誘発
CA3061984A1 (en) 2017-06-06 2018-12-13 Zymergen Inc. A htp genomic engineering platform for improving fungal strains
EP3635110A2 (en) 2017-06-06 2020-04-15 Zymergen, Inc. A high-throughput (htp) genomic engineering platform for improving saccharopolyspora spinosa
WO2018224544A1 (en) 2017-06-08 2018-12-13 Novozymes A/S Compositions comprising polypeptides having cellulase activity and amylase activity, and uses thereof in cleaning and detergent compositions
WO2018231462A1 (en) 2017-06-14 2018-12-20 Codexis, Inc. Engineered transaminase polypeptides for industrial biocatalysis
WO2018229715A1 (en) 2017-06-16 2018-12-20 Novartis Ag Compositions comprising anti-cd32b antibodies and methods of use thereof
CN111032871A (zh) 2017-06-20 2020-04-17 阿尔布梅迪克斯医疗有限公司 改良的表达蛋白质的菌株
WO2018234465A1 (en) 2017-06-22 2018-12-27 Novozymes A/S XYLANASE VARIANTS AND POLYNUCLEOTIDES ENCODING THE SAME
CN111051506B (zh) 2017-06-27 2023-11-24 科德克希思公司 青霉素g酰化酶
US11584783B2 (en) 2017-06-28 2023-02-21 Novozymes A/S Processes for producing a fermentation product
JP2020530267A (ja) 2017-06-30 2020-10-22 コデクシス， インコーポレイテッド Ｔ７ ｒｎａポリメラーゼバリアント
US10793841B2 (en) 2017-06-30 2020-10-06 Codexis, Inc. T7 RNA polymerase variants
CN111108183A (zh) 2017-06-30 2020-05-05 诺维信公司 酶浆液组合物
WO2019016772A2 (en) 2017-07-21 2019-01-24 Novartis Ag COMPOSITIONS AND METHODS FOR TREATING CANCER
US11746366B2 (en) 2017-07-24 2023-09-05 Novozymes A/S GH5 and GH30 in wet milling
CN111542608A (zh) 2017-07-28 2020-08-14 双刃基金会 马铃薯y病毒抗性基因及使用方法
EP4036105A3 (en) 2017-08-03 2022-11-02 AgBiome, Inc. Pesticidal genes and methods of use
WO2019030165A1 (en) 2017-08-08 2019-02-14 Novozymes A/S POLYPEPTIDES HAVING TREHALASE ACTIVITY AND THEIR USE IN A PRODUCTION PROCESS OF FERMENTATION PRODUCTS
US11634400B2 (en) 2017-08-19 2023-04-25 University Of Rochester Micheliolide derivatives, methods for their preparation and their use as anticancer and antiinflammatory agents
CN111344404A (zh) 2017-08-24 2020-06-26 诺维信公司 黄原胶裂解酶变体以及编码其的多核苷酸
EP3673060A1 (en) 2017-08-24 2020-07-01 Henkel AG & Co. KGaA Detergent composition comprising xanthan lyase variants ii
WO2019038059A1 (en) 2017-08-24 2019-02-28 Henkel Ag & Co. Kgaa DETERGENT COMPOSITIONS COMPRISING GH9 ENDOGLUCANASE VARIANTS II
EP3673058A1 (en) 2017-08-24 2020-07-01 Novozymes A/S Gh9 endoglucanase variants and polynucleotides encoding same
US11834484B2 (en) 2017-08-29 2023-12-05 Novozymes A/S Bakers's yeast expressing anti-staling/freshness amylases
CN111032879B (zh) 2017-08-31 2023-12-08 诺维信公司 具有d-阿洛酮糖3-差向异构酶活性的多肽和编码其的多核苷酸
EP3675646A1 (en) 2017-09-01 2020-07-08 Novozymes A/S Animal feed additives comprising polypeptide having protease activity and uses thereof
AR112955A1 (es) 2017-09-01 2020-01-08 Novozymes As Aditivos de alimento animal que comprenden polipéptidos que tienen actividad proteasa y usos de los mismos
WO2019046703A1 (en) 2017-09-01 2019-03-07 Novozymes A/S METHODS OF ENHANCING GENOME EDITION IN FUNGI
WO2019052907A2 (en) 2017-09-07 2019-03-21 Total Raffinage Chimie METHOD FOR DETECTING THE HYDROGEN PEROXIDE CONVERSION ACTIVITY OF AN ENZYME
WO2019047199A1 (en) 2017-09-11 2019-03-14 Danisco Us Inc. GLUCOAMYLASE AND METHODS OF USE THEREOF
WO2019060383A1 (en) 2017-09-25 2019-03-28 Pioneer Hi-Bred, International, Inc. PROMOTERS HAVING PREFERENCE FOR FABRICS AND METHODS OF USE
CN111108195A (zh) 2017-09-27 2020-05-05 宝洁公司 包含脂肪酶的洗涤剂组合物
CA3073362A1 (en) 2017-09-27 2019-04-04 Novozymes A/S Lipase variants and microcapsule compositions comprising such lipase variants
WO2019068713A1 (en) 2017-10-02 2019-04-11 Novozymes A/S POLYPEPTIDES HAVING MANNANASE ACTIVITY AND POLYNUCLEOTIDES ENCODING THESE POLYPEPTIDES
CN111373036A (zh) 2017-10-02 2020-07-03 诺维信公司 具有甘露聚糖酶活性的多肽和编码它们的多核苷酸
WO2019070883A1 (en) 2017-10-04 2019-04-11 Novozymes A/S POLYPEPTIDES WITH PROTEASE ACTIVITY AND POLYNUCLEOTIDES ENCODING SAME
US20200165626A1 (en) 2017-10-13 2020-05-28 Pioneer Hi-Bred International, Inc. Virus-induced gene silencing technology for insect control in maize
WO2019079629A1 (en) 2017-10-18 2019-04-25 Ascus Biosciences, Inc. IMPROVING POULTRY PRODUCTION BY ADMINISTERING A SYNTHETIC BIO-ASSEMBLY OF MICROBES OR PURIFIED STEM THEREFOR
WO2019083818A1 (en) 2017-10-23 2019-05-02 Novozymes A/S IMPROVING EXPRESSION OF PROTEASE BY CO-EXPRESSION WITH PROPEPTIDE
EP3700933A1 (en) 2017-10-25 2020-09-02 Novartis AG Antibodies targeting cd32b and methods of use thereof
MX2020004319A (es) 2017-10-25 2020-10-12 Da Volterra Variantes de beta-lactamasas.
CN117683748A (zh) 2017-10-27 2024-03-12 宝洁公司 包含多肽变体的洗涤剂组合物
EP3701016A1 (en) 2017-10-27 2020-09-02 Novozymes A/S Dnase variants
DE102017125558A1 (de) 2017-11-01 2019-05-02 Henkel Ag & Co. Kgaa Reinigungszusammensetzungen, die dispersine i enthalten
DE102017125559A1 (de) 2017-11-01 2019-05-02 Henkel Ag & Co. Kgaa Reinigungszusammensetzungen, die dispersine ii enthalten
DE102017125560A1 (de) 2017-11-01 2019-05-02 Henkel Ag & Co. Kgaa Reinigungszusammensetzungen, die dispersine iii enthalten
US11319531B2 (en) 2017-11-07 2022-05-03 Codexis, Inc. Transglutaminase variants
MX2020004863A (es) 2017-11-10 2020-10-14 Defensin Therapeutics Aps Maduracion de la defensa de la mucosa y/o funcion intestinal/pulmonar en el bebe prematuro.
US20200318086A1 (en) 2017-11-10 2020-10-08 Novozymes A/S Temperature-sensitive cas9 protein
BR112020009525A2 (pt) 2017-11-14 2020-11-03 Danisco Us Inc. alfa-amilase, composição e método
EP3954782A1 (en) 2017-11-15 2022-02-16 10X Genomics, Inc. Functionalized gel beads
US10829815B2 (en) 2017-11-17 2020-11-10 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
WO2019096903A1 (en) 2017-11-20 2019-05-23 Novozymes A/S New galactanases (ec 3.2.1.89) for use in soy processing
CN111741762A (zh) 2017-11-24 2020-10-02 防御素治疗学公司 使用防御素预防和治疗移植物抗宿主病
WO2019108619A1 (en) 2017-11-28 2019-06-06 Two Blades Foundation Methods and compositions for enhancing the disease resistance of plants
WO2019110462A1 (en) 2017-12-04 2019-06-13 Novozymes A/S Lipase variants and polynucleotides encoding same
WO2019113413A1 (en) 2017-12-08 2019-06-13 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
US11384347B2 (en) 2017-12-08 2022-07-12 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
JP7401435B2 (ja) 2017-12-13 2023-12-19 グラクソスミスクライン、インテレクチュアル、プロパティー、ディベロップメント、リミテッド カルボキシエステラーゼ生体触媒
CA3081950A1 (en) 2017-12-13 2019-06-20 Codexis, Inc. Carboxyesterase polypeptides for amide coupling
WO2019125651A1 (en) 2017-12-19 2019-06-27 Pioneer Hi-Bred International, Inc. Insecticidal polypeptides and uses thereof
CN111788306A (zh) 2017-12-20 2020-10-16 巴斯夫欧洲公司 核酸酶以及用于制备和使用其的方法
RU2020124098A (ru) 2017-12-22 2022-01-24 Новозимс А/С Способ помола пшеницы и ксиланазы gh8
BR112020012763A2 (pt) 2017-12-22 2020-12-15 AgBiome, Inc. Genes pesticidas e métodos de uso
MX2020007394A (es) 2018-01-10 2020-09-03 Brightseed Inc Metodo para modular el metabolismo.
WO2019140351A1 (en) 2018-01-12 2019-07-18 Two Blades Foundation Stem rust resistance genes and methods of use
MX2020007914A (es) 2018-01-29 2020-10-28 Novozymes As Microorganismos con uso de nitrogeno mejorado para produccion de etanol.
WO2019154951A1 (en) 2018-02-08 2019-08-15 Novozymes A/S Lipases, lipase variants and compositions thereof
US20210071156A1 (en) 2018-02-08 2021-03-11 Novozymes A/S Lipase Variants and Compositions Thereof
BR112020016306A2 (pt) 2018-02-12 2020-12-15 Curators Of The University Of Missouri Gene (saur) suprarregulado pequeno de auxina para o melhoramento da arquitetura do sistema radicular da planta, tolerância ao encharcamento, resistência à seca, e rendimento
CA3089284A1 (en) 2018-02-28 2019-09-06 The Procter & Gamble Company Methods of cleaning using a glycogen debranching enzyme
BR112020018294A2 (pt) 2018-03-09 2020-12-29 Danisco Us Inc. Glucoamilases e métodos de uso dos mesmos
CN111770788B (zh) 2018-03-13 2023-07-25 诺维信公司 使用氨基糖低聚物进行微囊化
CN111902540A (zh) 2018-03-20 2020-11-06 齐默尔根公司 用于中国仓鼠卵巢细胞基因工程改造的htp平台
US11359000B2 (en) 2018-03-28 2022-06-14 Bristol-Myers Squibb Company Interleukin-2/Interleukin-2 receptor alpha fusion proteins and methods of use
WO2019185726A1 (en) 2018-03-29 2019-10-03 Novozymes A/S Mannanase variants and polynucleotides encoding same
MX2020010092A (es) 2018-03-30 2020-10-28 Amgen Inc Variantes de anticuerpo c-terminales.
EP3775178A4 (en) 2018-04-06 2022-01-05 Braskem S.A. NEW NADH-DEPENDENT ENZYMMUTANTS FOR THE CONVERSION OF ACETONE INTO ISOPROPANOL
CN112262218A (zh) 2018-04-06 2021-01-22 10X基因组学有限公司 用于单细胞处理中的质量控制的系统和方法
BR112020020744A2 (pt) 2018-04-09 2021-02-02 Novozymes A/S polipeptídeos tendo atividade de alfa-amilase e polinucleotídeos codificando os mesmos
US11566239B2 (en) 2018-04-19 2023-01-31 Novozymes A/S Stabilized cellulase variants
US11661592B2 (en) 2018-04-19 2023-05-30 Novozymes A/S Stabilized endoglucanase variants
WO2019204584A1 (en) 2018-04-20 2019-10-24 AgBiome, Inc. Pesticidal proteins and methods of use
EP3794116A2 (en) 2018-05-17 2021-03-24 BP Corporation North America Inc. Production of 2-keto-3-deoxy-d-gluconic acid in filamentous fungi
US11702668B2 (en) 2018-05-22 2023-07-18 Pioneer Hi-Bred International, Inc. Plant regulatory elements and methods of use thereof
AR126019A1 (es) 2018-05-30 2023-09-06 Novartis Ag Anticuerpos frente a entpd2, terapias de combinación y métodos de uso de los anticuerpos y las terapias de combinación
BR112020022593A2 (pt) 2018-05-31 2021-02-09 Novozymes A/S método para tratar polpa de dissolução, polpa de dissolução, fibra têxtil ou celulose derivatizada, e, uso de uma monoxigenase lítica de polissacarídeo.
EP3802611A2 (en) 2018-06-01 2021-04-14 Novartis AG Binding molecules against bcma and uses thereof
SG11202011975WA (en) 2018-06-05 2020-12-30 Lifeedit Inc Rna-guided nucleases and active fragments and variants thereof and methods of use
KR20210018219A (ko) 2018-06-06 2021-02-17 지머젠 인코포레이티드 발효 및 생산 중 곰팡이 형태를 제어하기 위한 신호 전달에 포함된 유전자의 조작
WO2019241132A1 (en) 2018-06-12 2019-12-19 Codexis, Inc. Engineered tyrosine ammonia lyase
EP3814491A1 (en) 2018-06-27 2021-05-05 Boehringer Ingelheim RCV GmbH & Co KG Means and methods for increased protein expression by use of transcription factors
WO2020002575A1 (en) 2018-06-28 2020-01-02 Novozymes A/S Polypeptides having pectin lyase activity and polynucleotides encoding same
US20210277409A1 (en) 2018-06-28 2021-09-09 Pioneer Hi-Bred International, Inc. Methods for selecting transformed plants
WO2020010101A2 (en) 2018-07-04 2020-01-09 Danisco Us Inc Glucoamylases and methods of use, thereof
JP2021530981A (ja) 2018-07-09 2021-11-18 コデクシス， インコーポレイテッド 操作されたホスホペントムターゼ改変体酵素
EP3820502A4 (en) 2018-07-09 2022-07-20 Codexis, Inc. MODIFIED PURINE NUCLEOSIDE PHOSPHORYLASE ENZYME VARIANT
CA3103514A1 (en) 2018-07-09 2020-01-16 Codexis, Inc. Engineered galactose oxidase variant enzymes
SG11202012107RA (en) 2018-07-09 2021-01-28 Codexis Inc Engineered deoxyribose-phosphate aldolases
KR20210031929A (ko) 2018-07-09 2021-03-23 코덱시스, 인코포레이티드 조작된 판토테네이트 키나제 변이체 효소
WO2020014306A1 (en) 2018-07-10 2020-01-16 Immunogen, Inc. Met antibodies and immunoconjugates and uses thereof
WO2020013951A1 (en) 2018-07-12 2020-01-16 Codexis, Inc. Engineered phenylalanine ammonia lyase polypeptides
WO2020016459A1 (en) 2018-07-20 2020-01-23 Pierre Fabre Medicament Receptor for vista
CN112601818A (zh) 2018-07-25 2021-04-02 诺维信公司 用于生产乙醇的表达酶的酵母
BR112021001661A8 (pt) 2018-07-30 2022-08-09 Codexis Inc Glicosiltransferase modificada, polinucleotídeo modificado, vetor, célula hospedeira, métodos para produzir pelo menos uma glicosiltransferase modificada, para produzir pelo menos uma variante de sacarose sintase, para glicosilação de um substrato, para produzir rebaudiosídeo m, rebaudiosídeo a e/ou rebaudiosídeo i e rebaudiosídeo d, composição, sacarose sintase modificada, pelo menos um rebaudiosídeo, rebaudiosídeo m, rebaudiosídeo a, rebaudiosídeo i, e, rebaudiosídeo d.
US20210315984A1 (en) 2018-08-01 2021-10-14 Konstantin G. Kousoulas Compositions comprising herpes simplex virus-i for use in methods of treating and preventing cancer
MX2021002290A (es) 2018-08-29 2021-04-28 Pioneer Hi Bred Int Proteinas insecticidas y metodos para su uso.
WO2020043836A1 (en) 2018-08-31 2020-03-05 Novozymes A/S Polypeptides having protease activity and polynucleotides encoding same
CN112969792A (zh) 2018-09-07 2021-06-15 巴斯夫植物科学有限公司 用于在植物中产生高水平pufa的改进方法
CA3110656A1 (en) 2018-09-07 2020-03-12 Basf Plant Science Company Gmbh Improved method for the production of high levels of pufa in plants
CN113412332A (zh) 2018-09-07 2021-09-17 巴斯夫植物科学有限公司 用于在植物中产生高水平pufa的改进方法
US20220089977A1 (en) 2018-10-03 2022-03-24 Novozymes A/S Polypeptides Having Alpha-Mannan Degrading Activity And Polynucleotides Encoding Same
BR112021006692A2 (pt) 2018-10-08 2021-08-10 Novozymes A/S levedura expressando enzimas para produção de etanol
UY38407A (es) 2018-10-15 2020-05-29 Novartis Ag Anticuerpos estabilizadores de trem2
JP2022512847A (ja) 2018-10-29 2022-02-07 コデクシス， インコーポレイテッド 操作されたｄｎａポリメラーゼバリアント
CA3117805A1 (en) 2018-10-31 2020-05-07 Zymergen Inc. Multiplexed deterministic assembly of dna libraries
JP2022512817A (ja) 2018-10-31 2022-02-07 パイオニア ハイ－ブレッド インターナショナル， インコーポレイテッド オクロバクテリウム（Ｏｃｈｒｏｂａｃｔｒｕｍ）媒介植物形質転換のための組成物及び方法
CN113039278A (zh) 2018-10-31 2021-06-25 诺维信公司 通过指导的内切核酸酶和单链寡核苷酸进行基因组编辑
EP3873532A1 (en) 2018-10-31 2021-09-08 Novartis AG Dc-sign antibody drug conjugates
CN113302293A (zh) 2018-12-12 2021-08-24 诺维信公司 具有木聚糖酶活性的多肽以及编码这些多肽的多核苷酸
SG11202105668PA (en) 2018-12-14 2021-06-29 Codexis Inc Engineered tyrosine ammonia lyase
AU2019403323A1 (en) 2018-12-20 2021-07-01 Codexis, Inc. Human alpha-galactosidase variants
WO2020127796A2 (en) 2018-12-21 2020-06-25 Novozymes A/S Polypeptides having peptidoglycan degrading activity and polynucleotides encoding same
BR112021011900A2 (pt) 2018-12-21 2021-09-08 Novartis Ag Anticorpos para pmel17 e conjugados dos mesmos
US10973885B2 (en) 2018-12-27 2021-04-13 Colgate-Palmolive Company Oral care compositions
WO2020139783A2 (en) 2018-12-27 2020-07-02 Lifeedit, Inc. Polypeptides useful for gene editing and methods of use
EP3918060A1 (en) 2019-01-31 2021-12-08 Novozymes A/S Polypeptides having xylanase activity and use thereof for improving the nutritional quality of animal feed
SG11202107720UA (en) 2019-01-31 2021-08-30 Agency Science Tech & Res Cnx/erp57 inhibitor for use in the treatment or prevention of cancer
US11053515B2 (en) 2019-03-08 2021-07-06 Zymergen Inc. Pooled genome editing in microbes
CN113728106A (zh) 2019-03-08 2021-11-30 齐默尔根公司 微生物中的迭代基因组编辑
EP3938521A1 (en) 2019-03-11 2022-01-19 Pioneer Hi-Bred International, Inc. Methods for clonal plant production
AU2020242303A1 (en) 2019-03-21 2021-06-24 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
EP3947441A1 (en) 2019-03-27 2022-02-09 UMC Utrecht Holding B.V. Engineered iga antibodies and methods of use
WO2020198408A1 (en) 2019-03-27 2020-10-01 Pioneer Hi-Bred International, Inc. Plant explant transformation
WO2020198731A2 (en) 2019-03-28 2020-10-01 Danisco Us Inc Engineered antibodies
WO2020198496A1 (en) 2019-03-28 2020-10-01 Pioneer Hi-Bred International, Inc. Modified agrobacterium strains and use thereof for plant transformation
EP3947619A1 (en) 2019-04-03 2022-02-09 Novozymes A/S Polypeptides having beta-glucanase activity, polynucleotides encoding same and uses thereof in cleaning and detergent compositions
CN113874499A (zh) 2019-04-10 2021-12-31 诺维信公司 多肽变体
US20220186151A1 (en) 2019-04-12 2022-06-16 Novozymes A/S Stabilized glycoside hydrolase variants
WO2020229191A1 (en) 2019-05-15 2020-11-19 Novozymes A/S Temperature-sensitive rna-guided endonuclease
JP2022537887A (ja) 2019-05-21 2022-08-31 ノバルティス アーゲー Ｂｃｍａに対する三重特異性結合分子及びその使用
WO2020236797A1 (en) 2019-05-21 2020-11-26 Novartis Ag Variant cd58 domains and uses thereof
CR20210576A (es) 2019-05-21 2021-12-15 Novartis Ag Moléculas de unión a cd19 y usos de las mismas
CN113993996A (zh) 2019-06-24 2022-01-28 诺维信公司 α-淀粉酶变体
JP7326497B2 (ja) 2019-06-24 2023-08-15 ザ プロクター アンド ギャンブル カンパニー アミラーゼバリアントを含む洗浄組成物
WO2020264237A1 (en) 2019-06-27 2020-12-30 Two Blades Foundation Engineered atrlp23 pattern recognition receptors and methods of use
EP3994255A1 (en) 2019-07-02 2022-05-11 Novozymes A/S Lipase variants and compositions thereof
WO2021013464A1 (en) 2019-07-25 2021-01-28 Novozymes A/S Filamentous fungal expression system
EP4004279A1 (en) 2019-07-26 2022-06-01 Novozymes A/S Enzymatic treatment of paper pulp
BR112021026761A2 (pt) 2019-07-26 2022-02-15 Novozymes As Célula de levedura, composição, e, métodos de produção de um derivado de uma estirpe de levedura, de produção de etanol e de produção de um produto de fermentação a partir de um material contendo amido ou contendo celulose
KR102608787B1 (ko) 2019-07-29 2023-12-04 브라이트씨드, 인크. 소화기 건강의 개선 방법
CN114127124A (zh) 2019-08-06 2022-03-01 诺维信公司 用于提高酶表达的融合蛋白
AU2020329912A1 (en) 2019-08-12 2022-03-24 LifeEDIT Therapeutics, Inc. RNA-guided nucleases and active fragments and variants thereof and methods of use
JP2022545465A (ja) 2019-08-22 2022-10-27 ビーエーエスエフ ソシエタス・ヨーロピア アミラーゼバリアント
EP4031661A1 (en) 2019-09-16 2022-07-27 Novozymes A/S Polypeptides having beta-glucanase activity and polynucleotides encoding same
TW202124446A (zh) 2019-09-18 2021-07-01 瑞士商諾華公司 與ｅｎｔｐｄ２抗體之組合療法
EP4031578A1 (en) 2019-09-18 2022-07-27 Novartis AG Entpd2 antibodies, combination therapies, and methods of using the antibodies and combination therapies
WO2021067608A1 (en) 2019-10-02 2021-04-08 Abbott Laboratories Detection of analytes by protein switches
US20220340843A1 (en) 2019-10-03 2022-10-27 Novozymes A/S Polypeptides comprising at least two carbohydrate binding domains
KR20220100896A (ko) 2019-11-11 2022-07-18 브라이트씨드, 인크. 추출물, 소모가능 생성물 및 추출물에서 생물활성 대사산물을 농후화하는 방법
EP3821946A1 (en) 2019-11-12 2021-05-19 Université de Strasbourg Anti-claudin-1 monoclonal antibodies for the prevention and treatment of fibrotic diseases
US20230002794A1 (en) 2019-12-10 2023-01-05 Novozymes A/S Microorganism for improved pentose fermentation
CA3160401A1 (en) 2019-12-19 2021-06-24 Neil Joseph Lant Cleaning compositions comprising polypeptides having alpha amylase activity
MX2022007450A (es) 2019-12-19 2022-06-27 Novozymes As Variantes de xilanasa y polinucleotidos que codifican las mismas.
BR112022011955A2 (pt) 2019-12-19 2022-09-06 Novozymes As Variantes de alfa-amilase
CA3161898A1 (en) 2019-12-19 2021-06-24 Basf Se Increasing space-time-yield, carbon-conversion-efficiency and carbon substrate flexibility in the production of fine chemicals
WO2021123307A2 (en) 2019-12-20 2021-06-24 Novozymes A/S Polypeptides having proteolytic activity and use thereof
BR112022011895A2 (pt) 2019-12-20 2022-09-06 Basf Se Diminuição da toxidez de terpenos e aumento da produção potencial em microrganismos
CN115190912A (zh) 2019-12-30 2022-10-14 生命编辑制药股份有限公司 Rna指导核酸酶及其活性片段与变体以及使用方法
BR112022014573A2 (pt) 2020-01-24 2022-10-04 Novozymes As Mutantes de uma célula fúngica filamentosa com produtividade aumentada na produção de um polipeptídeo
WO2021152120A1 (en) 2020-01-31 2021-08-05 Novozymes A/S Mannanase variants and polynucleotides encoding same
CN115052981A (zh) 2020-01-31 2022-09-13 诺维信公司 甘露聚糖酶变体以及编码它们的多核苷酸
US20230142991A1 (en) 2020-02-10 2023-05-11 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
WO2021163030A2 (en) 2020-02-10 2021-08-19 Novozymes A/S Polypeptides having alpha-amylase activity and polynucleotides encoding same
US20230235367A1 (en) 2020-02-10 2023-07-27 Novozymes A/S Process for producing ethanol from raw starch using alpha-amylase variants
CN115349128A (zh) 2020-02-13 2022-11-15 齐默尔根公司 宏基因组文库和天然产物发现平台
US20230090771A1 (en) * 2020-02-21 2023-03-23 Regents Of The University Of Minnesota Novel methods for creating alpha-n-methylated polypeptides
US20230220366A1 (en) 2020-02-26 2023-07-13 Novozymes A/S Polypeptide Variants
WO2021202473A2 (en) 2020-03-30 2021-10-07 Danisco Us Inc Engineered antibodies
WO2021202463A1 (en) 2020-03-30 2021-10-07 Danisco Us Inc Anti-rsv antibodies
WO2021204838A1 (en) 2020-04-08 2021-10-14 Novozymes A/S Carbohydrate binding module variants
WO2021207200A2 (en) 2020-04-10 2021-10-14 Codexis, Inc. Engineered transaminase polypeptides
CA3175633A1 (en) 2020-04-17 2021-10-21 Danisco Us Inc. Glucoamylase and methods of use thereof
EP4139431A1 (en) 2020-04-21 2023-03-01 Novozymes A/S Cleaning compositions comprising polypeptides having fructan degrading activity
TW202208626A (zh) 2020-04-24 2022-03-01 美商生命編輯公司 Rna引導核酸酶及其活性片段與變體，以及使用方法
US20230181756A1 (en) 2020-04-30 2023-06-15 Novartis Ag Ccr7 antibody drug conjugates for treating cancer
CN116096758A (zh) 2020-05-01 2023-05-09 诺华股份有限公司 工程化免疫球蛋白
US20230167193A1 (en) 2020-05-01 2023-06-01 Novartis Ag Immunoglobulin variants
WO2021231437A1 (en) 2020-05-11 2021-11-18 LifeEDIT Therapeutics, Inc. Rna-guided nucleic acid binding proteins and active fragments and variants thereof and methods of use
WO2021245064A1 (en) 2020-06-04 2021-12-09 Isobionics B.V. Synthetic santalene synthases
WO2022003142A1 (en) 2020-07-03 2022-01-06 Engenes Biotech Gmbh PYRROLYSYL-tRNA SYNTHETASE VARIANTS AND USES THEREOF
EP4182466A2 (en) 2020-07-14 2023-05-24 Pioneer Hi-Bred International, Inc. Insecticidal proteins and methods for their use
IL299812A (en) 2020-07-15 2023-03-01 Lifeedit Therapeutics Inc Uracil stabilizing proteins and active fragments and their variants and methods of use
US11479779B2 (en) 2020-07-31 2022-10-25 Zymergen Inc. Systems and methods for high-throughput automated strain generation for non-sporulating fungi
WO2022035653A2 (en) 2020-08-10 2022-02-17 E. I. Du Pont De Nemours And Company Compositions and methods for enhancing resistance to northern leaf blight in maize
EP4192848A1 (en) 2020-08-10 2023-06-14 Pioneer Hi-Bred International, Inc. Plant regulatory elements and methods of use thereof
EP4196575A1 (en) 2020-08-13 2023-06-21 Novozymes A/S Phytase variants and polynucleotides encoding same
US20230313209A1 (en) 2020-08-18 2023-10-05 Novozymes A/S Dispersins expressed with amylase signal peptides
JP2023538740A (ja) 2020-08-25 2023-09-11 ノボザイムス アクティーゼルスカブ ファミリー４４キシログルカナーゼの変異体
EP4204547A1 (en) 2020-08-28 2023-07-05 Novozymes A/S Protease variants with improved solubility
US11767519B2 (en) 2020-08-28 2023-09-26 Codexis, Inc. Engineered amylase variants
EP4211235A2 (en) 2020-09-11 2023-07-19 Lifeedit Therapeutics, Inc. Dna modifying enzymes and active fragments and variants thereof and methods of use
EP4210758A1 (en) 2020-09-14 2023-07-19 Aesculus Bio ApS Defensin fragment derived lipopeptides for treatment of drug-resistant microorganisms
JP2023544016A (ja) 2020-09-30 2023-10-19 パイオニア ハイ－ブレッド インターナショナル， インコーポレイテッド 単子葉外植片の迅速な形質転換
CN116507725A (zh) 2020-10-07 2023-07-28 诺维信公司 α-淀粉酶变体
EP4229189A1 (en) 2020-10-13 2023-08-23 Novozymes A/S Glycosyltransferase variants for improved protein production
CN116615523A (zh) 2020-10-28 2023-08-18 诺维信公司 脂氧合酶的用途
WO2022090361A2 (en) 2020-10-29 2022-05-05 Novozymes A/S Lipase variants and compositions comprising such lipase variants
MX2023004938A (es) 2020-11-02 2023-05-17 Novozymes As Variantes de glucoamilasa y polinucleotidos que codifican las mismas.
WO2022090555A1 (en) 2020-11-02 2022-05-05 Novozymes A/S Leader peptides and polynucleotides encoding the same
US20240025993A1 (en) 2020-11-06 2024-01-25 Novartis Ag Cd19 binding molecules and uses thereof
WO2022097061A1 (en) 2020-11-06 2022-05-12 Novartis Ag Anti-cd19 agent and b cell targeting agent combination therapy for treating b cell malignancies
WO2022106404A1 (en) 2020-11-18 2022-05-27 Novozymes A/S Combination of proteases
WO2022106400A1 (en) 2020-11-18 2022-05-27 Novozymes A/S Combination of immunochemically different proteases
JP2023549925A (ja) 2020-11-24 2023-11-29 ノバルティス アーゲー 抗ｃｄ４８抗体、抗体薬物コンジュゲート及びその使用
EP4251755A2 (en) 2020-11-24 2023-10-04 AgBiome, Inc. Pesticidal genes and methods of use
CA3201757A1 (en) 2020-12-11 2022-06-16 Mathias Schuetz Recombinant acyl activating enzyme (aae) genes for enhanced biosynthesis of cannabinoids and cannabinoid precursors
CN116710471A (zh) 2020-12-15 2023-09-05 诺维信公司 具有减少的细胞运动的突变的宿主细胞
EP4015626A1 (en) 2020-12-18 2022-06-22 Isobionics B.V. Enzymes and methods for fermentative production of monoterpene esters
JP2024500102A (ja) 2020-12-18 2024-01-04 コデクシス， インコーポレイテッド 操作されたウリジンホスホリラーゼ改変体酵素
EP4271482A2 (en) 2020-12-31 2023-11-08 Alamar Biosciences, Inc. Binder molecules with high affinity and/ or specificity and methods of making and use thereof
CN112725331B (zh) * 2021-01-25 2021-07-20 深圳市狂风生命科技有限公司 一种高通量突变体文库的构建方法
CN116829685A (zh) 2021-01-28 2023-09-29 诺维信公司 具有低恶臭产生的脂肪酶
EP4291646A2 (en) 2021-02-12 2023-12-20 Novozymes A/S Alpha-amylase variants
AR124921A1 (es) 2021-02-18 2023-05-17 Novozymes As Polipéptidos inactivos que contienen hemo
EP4047088A1 (en) 2021-02-22 2022-08-24 Basf Se Amylase variants
WO2022175435A1 (en) 2021-02-22 2022-08-25 Basf Se Amylase variants
EP4305146A1 (en) 2021-03-12 2024-01-17 Novozymes A/S Polypeptide variants
EP4060036A1 (en) 2021-03-15 2022-09-21 Novozymes A/S Polypeptide variants
US20240060061A1 (en) 2021-03-15 2024-02-22 Novozymes A/S Dnase variants
JP2023548846A (ja) 2021-03-15 2023-11-21 ザ プロクター アンド ギャンブル カンパニー ポリペプチドバリアントを含有する洗浄組成物
TW202300649A (zh) 2021-03-22 2023-01-01 美商生命編輯治療學公司 Ｄｎａ修飾酶及活性片段及其變體及使用方法
CN117083370A (zh) 2021-03-26 2023-11-17 诺维信公司 聚合物含量降低的洗涤剂组合物
EP4314264A1 (en) 2021-04-02 2024-02-07 Codexis, Inc. Engineered cyclic gmp-amp synthase (cgas) variant enzymes
US11913037B2 (en) 2021-04-02 2024-02-27 Codexis, Inc. Engineered adenylate kinase variant enzymes
CN117222735A (zh) 2021-04-02 2023-12-12 科德克希思公司 工程化乙酸激酶变体酶
EP4314262A1 (en) 2021-04-02 2024-02-07 Codexis, Inc. Engineered guanylate kinase variant enzymes
CN117529235A (zh) 2021-04-12 2024-02-06 诺维信公司 用于生产人造肉产品的方法
WO2022221720A1 (en) 2021-04-16 2022-10-20 Novartis Ag Antibody drug conjugates and methods for making thereof
AR125794A1 (es) 2021-05-06 2023-08-16 Agbiome Inc Genes plaguicidas y métodos de uso
CA3219611A1 (en) 2021-05-11 2022-11-17 Two Blades Foundation Methods for preparing a library of plant disease resistance genes for functional testing for disease resistance
BR112023024706A2 (pt) 2021-05-27 2024-02-15 Novozymes As Reguladores da transcrição e polinucleotídeos que os codificam
CN117795089A (zh) 2021-06-07 2024-03-29 诺维信公司 用于改善的乙醇发酵的工程化微生物
CA3173953A1 (en) 2021-06-11 2023-12-10 Tyson D. BOWEN Rna polymerase iii promoters and methods of use
CA3220135A1 (en) 2021-06-16 2022-12-22 Novozymes A/S Method for controlling slime in a pulp or paper making process
WO2023285348A1 (en) 2021-07-13 2023-01-19 Novozymes A/S Recombinant cutinase expression
WO2023288294A1 (en) 2021-07-16 2023-01-19 Novozymes A/S Compositions and methods for improving the rainfastness of proteins on plant surfaces
WO2023010083A2 (en) 2021-07-30 2023-02-02 Willow Biosciences, Inc. Recombinant prenyltransferase polypeptides engineered for enhanced biosynthesis of cannabinoids
CN117795087A (zh) 2021-08-02 2024-03-29 巴斯夫欧洲公司 用紫罗兰亚基乙烷合酶进行的芳香化合物的新颖生产
CA3227236A1 (en) 2021-08-19 2023-02-23 Trish Choudhary Recombinant olivetolic acid cyclase polypeptides engineered for enhanced biosynthesis of cannabinoids
WO2023056269A1 (en) 2021-09-30 2023-04-06 Two Blades Foundation Plant disease resistance genes against stem rust and methods of use
WO2023061928A1 (en) 2021-10-12 2023-04-20 Novozymes A/S Endoglucanase with improved stability
WO2023064927A1 (en) 2021-10-15 2023-04-20 Cytomx Therapeutics, Inc. Activatable polypeptide complex
WO2023064905A1 (en) 2021-10-15 2023-04-20 Danisco Us Inc. Glucoamylase variants and methods for use thereof
WO2023064929A1 (en) 2021-10-15 2023-04-20 Cytomx Therapeutics, Inc. Activatable polypeptide complex
WO2023064955A1 (en) 2021-10-15 2023-04-20 Cytomx Therapeutics, Inc. Activatable anti-cd3, anti-egfr, heteromultimeric bispecific polypeptide complex
WO2023069921A1 (en) 2021-10-19 2023-04-27 Epimeron Usa, Inc. Recombinant thca synthase polypeptides engineered for enhanced biosynthesis of cannabinoids
WO2023107943A1 (en) 2021-12-07 2023-06-15 AgBiome, Inc. Pesticidal genes and methods of use
WO2023104846A1 (en) 2021-12-10 2023-06-15 Novozymes A/S Improved protein production in recombinant bacteria
WO2023118436A1 (en) 2021-12-22 2023-06-29 Novozymes A/S Method of producing a milk-based product
WO2023118565A1 (en) 2021-12-23 2023-06-29 Novozymes A/S Reduction of residual dna in microbial fermentation products
WO2023131901A1 (en) 2022-01-07 2023-07-13 Johnson & Johnson Enterprise Innovation Inc. Materials and methods of il-1beta binding proteins
WO2023139557A1 (en) 2022-01-24 2023-07-27 LifeEDIT Therapeutics, Inc. Rna-guided nucleases and active fragments and variants thereof and methods of use
WO2023152220A1 (en) 2022-02-10 2023-08-17 Novozymes A/S Improved expression of recombinant proteins
WO2023161245A1 (en) 2022-02-22 2023-08-31 Novozymes A/S Oral care composition comprising enzymes
WO2023165507A1 (en) 2022-03-02 2023-09-07 Novozymes A/S Use of xyloglucanase for improvement of sustainability of detergents
WO2023170177A1 (en) 2022-03-08 2023-09-14 Novozymes A/S Fusion polypeptides with deamidase inhibitor and deamidase domains
EP4242303A1 (en) 2022-03-08 2023-09-13 Novozymes A/S Fusion polypeptides with deamidase inhibitor and deamidase domains
WO2023173084A1 (en) 2022-03-11 2023-09-14 University Of Rochester Cyclopeptibodies and uses thereof
TW202400658A (zh) 2022-04-26 2024-01-01 瑞士商諾華公司 靶向ｉｌ—１３和ｉｌ—１８的多特異性抗體
WO2023225459A2 (en) 2022-05-14 2023-11-23 Novozymes A/S Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections
WO2023222648A2 (en) 2022-05-17 2023-11-23 Novozymes A/S Process for reducing free fatty acids
WO2023247348A1 (en) 2022-06-21 2023-12-28 Novozymes A/S Mannanase variants and polynucleotides encoding same
WO2023247514A1 (en) 2022-06-22 2023-12-28 Novozymes A/S Recombinant mannanase expression
WO2023247664A2 (en) 2022-06-24 2023-12-28 Novozymes A/S Lipase variants and compositions comprising such lipase variants
WO2024003143A1 (en) 2022-06-30 2024-01-04 Novozymes A/S Mutanases and oral care compositions comprising same
WO2024013727A1 (en) 2022-07-15 2024-01-18 Janssen Biotech, Inc. Material and methods for improved bioengineered pairing of antigen-binding variable regions
WO2024012912A1 (en) 2022-07-15 2024-01-18 Novozymes A/S Polypeptides having deamidase inhibitor activity
WO2024015953A1 (en) 2022-07-15 2024-01-18 Danisco Us Inc. Methods for producing monoclonal antibodies
EP4311430A1 (en) 2022-07-28 2024-01-31 Limagrain Europe Chlorotoluron tolerance gene and methods of use thereof
WO2024026406A2 (en) 2022-07-29 2024-02-01 Vestaron Corporation Next Generation ACTX Peptides
WO2024033134A1 (en) 2022-08-11 2024-02-15 Basf Se Enzyme compositions comprising protease, mannanase, and/or cellulase
WO2024033133A2 (en) 2022-08-11 2024-02-15 Basf Se Enzyme compositions comprising an amylase
WO2024033901A1 (en) 2022-08-12 2024-02-15 LifeEDIT Therapeutics, Inc. Rna-guided nucleases and active fragments and variants thereof and methods of use
WO2024040020A1 (en) 2022-08-15 2024-02-22 Absci Corporation Quantitative affinity activity specific cell enrichment
WO2024044596A1 (en) 2022-08-23 2024-02-29 AgBiome, Inc. Pesticidal genes and methods of use
WO2024042489A1 (en) 2022-08-25 2024-02-29 LifeEDIT Therapeutics, Inc. Chemical modification of guide rnas with locked nucleic acid for rna guided nuclease-mediated gene editing
WO2024056643A1 (en) 2022-09-15 2024-03-21 Novozymes A/S Fungal signal peptides
EP4273249A2 (en) 2023-07-07 2023-11-08 Novozymes A/S Improved expression of recombinant proteins

Family Cites Families (154)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4994379A (en) * 1983-03-09 1991-02-19 Cetus Corporation Modified signal peptides
US4816567A (en) * 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US5422266A (en) 1984-12-31 1995-06-06 University Of Georgia Research Foundation, Inc. Recombinant DNA vectors capable of expressing apoaequorin
US4965188A (en) * 1986-08-22 1990-10-23 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US5176995A (en) * 1985-03-28 1993-01-05 Hoffmann-La Roche Inc. Detection of viruses by amplification and hybridization
CH0229046H1 (de) 1985-03-30 1998-07-15 Stuart Alan Kauffman Method for obtaining dna, rna, peptides, polypeptinique. des or proteins by means of a dna recombinant tech
US4959312A (en) 1985-05-31 1990-09-25 The University Of Tennessee Research Corporation Full spectrum mutagenesis
EP0208491B1 (en) * 1985-07-03 1993-08-25 Genencor International, Inc. Hybrid polypeptides and process for their preparation
US5866363A (en) 1985-08-28 1999-02-02 Pieczenik; George Method and means for sorting and identifying biological information
US4800159A (en) * 1986-02-07 1989-01-24 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences
DK311186D0 (da) * 1986-06-30 1986-06-30 Novo Industri As Enzymer
US5824469A (en) * 1986-07-17 1998-10-20 University Of Washington Method for producing novel DNA sequences with biological activity
US5418149A (en) 1990-07-24 1995-05-23 Hoffmann-La Roche Inc. Reduction of non-specific amplification glycosylase using DUTP and DNA uracil
US5763192A (en) 1986-11-20 1998-06-09 Ixsys, Incorporated Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique
US4994368A (en) * 1987-07-23 1991-02-19 Syntex (U.S.A.) Inc. Amplification method for polynucleotide assays
US4994367A (en) 1988-10-07 1991-02-19 East Carolina University Extended shelf life platelet preparations and process for preparing the same
FR2641793B1 (fr) * 1988-12-26 1993-10-01 Setratech Procede de recombinaison in vivo de sequences d'adn presentant des mesappariements de bases
DD282028A5 (de) 1989-02-16 1990-08-29 Akad Wissenschaften Ddr Verfahren zur herstellung einer thermostabilen, hybriden bacillus-beta-1,3-1,4-glukanase
US5106727A (en) * 1989-04-27 1992-04-21 Life Technologies, Inc. Amplification of nucleic acid sequences using oligonucleotides of random sequences as primers
US5043272A (en) * 1989-04-27 1991-08-27 Life Technologies, Incorporated Amplification of nucleic acid sequences using oligonucleotides of random sequence as primers
US5556750A (en) * 1989-05-12 1996-09-17 Duke University Methods and kits for fractionating a population of DNA molecules based on the presence or absence of a base-pair mismatch utilizing mismatch repair systems
CA2016841C (en) * 1989-05-16 1999-09-21 William D. Huse A method for producing polymers having a preselected activity
CA2016842A1 (en) * 1989-05-16 1990-11-16 Richard A. Lerner Method for tapping the immunological repertoire
IT1231157B (it) * 1989-07-20 1991-11-19 Crc Ricerca Chim Nuovi geni ibridi funzionali di bacillus thuringiensis ottenuti mediante ricombinazione in vivo.
US5574205A (en) 1989-07-25 1996-11-12 Cell Genesys Homologous recombination for universal donor cells and chimeric mammalian hosts
US5023171A (en) * 1989-08-10 1991-06-11 Mayo Foundation For Medical Education And Research Method for gene splicing by overlap extension using the polymerase chain reaction
WO1991006570A1 (en) * 1989-10-25 1991-05-16 The University Of Melbourne HYBRID Fc RECEPTOR MOLECULES
US5071743A (en) 1989-10-27 1991-12-10 Her Majesty The Queen In Right Of Canada, As Represented By The National Research Council Of Canada Process for conducting site-directed mutagenesis
DE3936258C1 (zh) 1989-10-31 1991-04-25 Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften Ev, 3400 Goettingen, De
WO1991007506A1 (en) * 1989-11-08 1991-05-30 The United States Of America, Represented By The Secretary, United States Department Of Commerce A method of synthesizing double-stranded dna molecules
EP0527809B1 (en) * 1990-04-05 1995-08-16 CREA, Roberto Walk-through mutagenesis
EP0528881A4 (en) 1990-04-24 1993-05-26 Stratagene Methods for phenotype creation from multiple gene populations
US5877402A (en) 1990-05-01 1999-03-02 Rutgers, The State University Of New Jersey DNA constructs and methods for stably transforming plastids of multicellular plants and expressing recombinant proteins therein
US5851813A (en) * 1990-07-12 1998-12-22 President And Fellows Of Harvard College Primate lentivirus antigenic compositions
US5571208A (en) 1990-07-13 1996-11-05 Caspers; Carl A. Reinforced prosthetic polyurethane hypobaric sleeve
US5169764A (en) 1990-08-08 1992-12-08 Regeneron Pharmaceuticals, Inc. Multitrophic and multifunctional chimeric neurotrophic factors, and nucleic acids and plasmids encoding the chimeras
WO1992003461A1 (en) * 1990-08-24 1992-03-05 Ixsys, Inc. Methods of synthesizing oligonucleotides with random codons
US5264563A (en) * 1990-08-24 1993-11-23 Ixsys Inc. Process for synthesizing oligonucleotides with random codons
US5698426A (en) * 1990-09-28 1997-12-16 Ixsys, Incorporated Surface expression libraries of heteromeric receptors
US5871974A (en) * 1990-09-28 1999-02-16 Ixsys Inc. Surface expression libraries of heteromeric receptors
US5770434A (en) * 1990-09-28 1998-06-23 Ixsys Incorporated Soluble peptides having constrained, secondary conformation in solution and method of making same
IL99553A0 (en) * 1990-09-28 1992-08-18 Ixsys Inc Compositions containing oligonucleotides linked to expression elements,a kit for the preparation of vectors useful for the expression of a diverse population of random peptides and methods utilizing the same
US5858725A (en) * 1990-10-10 1999-01-12 Glaxo Wellcome Inc. Preparation of chimaeric antibodies using the recombinant PCR strategy
US5234824A (en) * 1990-11-13 1993-08-10 Specialty Laboratories, Inc. Rapid purification of DNA
US5187083A (en) * 1990-11-13 1993-02-16 Specialty Laboratories, Inc. Rapid purification of DNA
US5489523A (en) * 1990-12-03 1996-02-06 Stratagene Exonuclease-deficient thermostable Pyrococcus furiosus DNA polymerase I
WO1992011272A1 (en) * 1990-12-20 1992-07-09 Ixsys, Inc. Optimization of binding proteins
DE4112440C1 (zh) * 1991-04-16 1992-10-22 Diagen Institut Fuer Molekularbiologische Diagnostik Gmbh, 4000 Duesseldorf, De
US5795747A (en) * 1991-04-16 1998-08-18 Evotec Biosystems Gmbh Process for manufacturing new biopolymers
JPH06510116A (ja) * 1991-04-17 1994-11-10 ベミス・マニファクチュアリング・カンパニー データ点解析の方法及び装置
US5512463A (en) * 1991-04-26 1996-04-30 Eli Lilly And Company Enzymatic inverse polymerase chain reaction library mutagenesis
US5514568A (en) * 1991-04-26 1996-05-07 Eli Lilly And Company Enzymatic inverse polymerase chain reaction
EP0519338B1 (en) 1991-06-20 1996-08-28 F. Hoffmann-La Roche Ag Improved methods for nucleic acid amplification
WO1993000103A1 (en) * 1991-06-21 1993-01-07 The Wistar Institute Of Anatomy And Biology Chimeric envelope proteins for viral targeting
EP0592597B1 (en) 1991-07-01 2001-09-26 Berlex Laboratories, Inc. Novel mutagenesis methods and compositions
US6071889A (en) 1991-07-08 2000-06-06 Neurospheres Holdings Ltd. In vivo genetic modification of growth factor-responsive neural precursor cells
US5223408A (en) * 1991-07-11 1993-06-29 Genentech, Inc. Method for making variant secreted proteins with altered properties
GB9115364D0 (en) * 1991-07-16 1991-08-28 Wellcome Found Antibody
US5279952A (en) * 1991-08-09 1994-01-18 Board Of Regents, The University Of Texas System PCR-based strategy of constructing chimeric DNA molecules
WO1993006213A1 (en) * 1991-09-23 1993-04-01 Medical Research Council Production of chimeric antibodies - a combinatorial approach
GB9125979D0 (en) * 1991-12-06 1992-02-05 Wellcome Found Antibody
FR2685004A1 (fr) * 1991-12-11 1993-06-18 Pasteur Institut Procede de generation d'une diversite structurale et fonctionnelle dans une sequence peptidique.
JP3431146B2 (ja) * 1992-01-30 2003-07-28 ジェンザイム・リミテッド 改変した酵素によるキラル合成
US5773267A (en) 1992-02-07 1998-06-30 Albert Einstein College Of Medicine Of Yeshiva University, A Division Of Yeshiva University D29 shuttle phasmids and uses thereof
GB9202796D0 (en) * 1992-02-11 1992-03-25 Wellcome Found Antiviral antibody
US5843643A (en) 1992-02-21 1998-12-01 Ratner; Paul L. Site-specific transfection of eukaryotic cells using polypeptide-linked recombinant nucleic acid
JPH07507995A (ja) * 1992-03-02 1995-09-07 バイオジェン，インコーポレイテッド トロンビンレセプターアンタゴニスト
WO1993019170A1 (en) 1992-03-16 1993-09-30 Jacob Nathaniel Wohlstadter Selection methods
WO1993019172A1 (en) 1992-03-24 1993-09-30 Cambridge Antibody Technology Limited Methods for producing members of specific binding pairs
US5316935A (en) 1992-04-06 1994-05-31 California Institute Of Technology Subtilisin variants suitable for hydrolysis and synthesis in organic media
DK0786257T3 (da) 1992-06-03 2003-10-06 Genentech Inc Glycolyseringsvarianter af vævsplasminogenaktivator med forbedrede terapeutiske egenskaber
EP0606430A1 (en) * 1992-06-15 1994-07-20 City Of Hope Chimeric anti-cea antibody
MX9304135A (es) * 1992-07-10 1995-01-31 Energy Biosystems Corp Adn recombinante que codifica un biocatalizador de desulfuracion.
US6107062A (en) * 1992-07-30 2000-08-22 Inpax, Inc. Antisense viruses and antisense-ribozyme viruses
FR2694754B1 (fr) 1992-08-12 1994-09-16 Bio Merieux Fragments d'ADN de mycobactéries, amorces d'amplification, sondes d'hybridation, réactifs et procédé de détection de détection de mycobactéries.
DE69232604T2 (de) * 1992-11-04 2002-11-07 City Of Hope Duarte Antikörperkonstrukte
WO1994011496A1 (en) * 1992-11-10 1994-05-26 Ixsys, Inc. Soluble peptides having constrained, secondary conformation in solution and method of making same
US5360728A (en) * 1992-12-01 1994-11-01 Woods Hole Oceanographic Institution (W.H.O.I.) Modified apoaequorin having increased bioluminescent activity
ATE199392T1 (de) * 1992-12-04 2001-03-15 Medical Res Council Multivalente und multispezifische bindungsproteine, deren herstellung und verwendung
US5571708A (en) 1993-04-19 1996-11-05 Bristol-Myers Squibb Company Thrombin-activatable plasminogen activator
IT1264712B1 (it) * 1993-07-13 1996-10-04 Eniricerche Spa Metodo di sintesi biologica di peptidi
US5498531A (en) 1993-09-10 1996-03-12 President And Fellows Of Harvard College Intron-mediated recombinant techniques and reagents
US5474920A (en) 1993-11-23 1995-12-12 State Of Oregon, Acting By And Through The Oregon State Board Of Higher Education On Behalf Of The Oregon Health Sciences University Modified thermo-resistant DNA polymerases
US5556772A (en) * 1993-12-08 1996-09-17 Stratagene Polymerase compositions and uses thereof
DE4343591A1 (de) * 1993-12-21 1995-06-22 Evotec Biosystems Gmbh Verfahren zum evolutiven Design und Synthese funktionaler Polymere auf der Basis von Formenelementen und Formencodes
US6335160B1 (en) * 1995-02-17 2002-01-01 Maxygen, Inc. Methods and compositions for polypeptide engineering
US5834252A (en) 1995-04-18 1998-11-10 Glaxo Group Limited End-complementary polymerase reaction
US5605793A (en) * 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination
US6117679A (en) 1994-02-17 2000-09-12 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US5837458A (en) 1994-02-17 1998-11-17 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US5928905A (en) * 1995-04-18 1999-07-27 Glaxo Group Limited End-complementary polymerase reaction
US6395547B1 (en) * 1994-02-17 2002-05-28 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6995017B1 (en) * 1994-02-17 2006-02-07 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6165793A (en) * 1996-03-25 2000-12-26 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US5521077A (en) * 1994-04-28 1996-05-28 The Leland Stanford Junior University Method of generating multiple protein variants and populations of protein variants prepared thereby
WO1996017056A1 (en) 1994-12-02 1996-06-06 Institut Pasteur Hypermutagenesis
EP0714980A1 (en) 1994-12-02 1996-06-05 Institut Pasteur Hypermutagenesis
US5514588A (en) 1994-12-13 1996-05-07 Exxon Research And Engineering Company Surfactant-nutrients for bioremediation of hydrocarbon contaminated soils and water
US5629179A (en) * 1995-03-17 1997-05-13 Novagen, Inc. Method and kit for making CDNA library
EP0822990A4 (en) * 1995-04-24 2002-07-03 Chromaxome Corp METHODS OF GENERATING AND SCREENING NEW METABOLIC PATHWAYS
US6030779A (en) 1995-07-18 2000-02-29 Diversa Corporation Screening for novel bioactivities
US6168919B1 (en) 1996-07-17 2001-01-02 Diversa Corporation Screening methods for enzymes and enzyme kits
US5958672A (en) * 1995-07-18 1999-09-28 Diversa Corporation Protein activity screening of clones having DNA from uncultivated microorganisms
US6057103A (en) 1995-07-18 2000-05-02 Diversa Corporation Screening for novel bioactivities
US6004788A (en) 1995-07-18 1999-12-21 Diversa Corporation Enzyme kits and libraries
JP4156666B2 (ja) * 1995-08-11 2008-09-24 ノボザイムス アクティーゼルスカブ ポリペプチド変異体を調整するための方法
US5962258A (en) 1995-08-23 1999-10-05 Diversa Corporation Carboxymethyl cellulase fromthermotoga maritima
FR2738840B1 (fr) 1995-09-19 1997-10-31 Rhone Poulenc Rorer Sa Souches de levures genetiquement modifiees
US6051409A (en) 1995-09-25 2000-04-18 Novartis Finance Corporation Method for achieving integration of exogenous DNA delivered by non-biological means to plant cells
US5756316A (en) * 1995-11-02 1998-05-26 Genencor International, Inc. Molecular cloning by multimerization of plasmids
US6171820B1 (en) 1995-12-07 2001-01-09 Diversa Corporation Saturation mutagenesis in directed evolution
US6238884B1 (en) 1995-12-07 2001-05-29 Diversa Corporation End selection in directed evolution
US6358709B1 (en) 1995-12-07 2002-03-19 Diversa Corporation End selection in directed evolution
US20030215798A1 (en) 1997-06-16 2003-11-20 Diversa Corporation High throughput fluorescence-based screening for novel enzymes
US5965408A (en) * 1996-07-09 1999-10-12 Diversa Corporation Method of DNA reassembly by interrupting synthesis
US6352842B1 (en) 1995-12-07 2002-03-05 Diversa Corporation Exonucease-mediated gene assembly in directed evolution
US6361974B1 (en) 1995-12-07 2002-03-26 Diversa Corporation Exonuclease-mediated nucleic acid reassembly in directed evolution
US5962283A (en) 1995-12-07 1999-10-05 Diversa Corporation Transminases and amnotransferases
US5814473A (en) 1996-02-09 1998-09-29 Diversa Corporation Transaminases and aminotransferases
US5830696A (en) * 1996-12-05 1998-11-03 Diversa Corporation Directed evolution of thermophilic enzymes
US5939250A (en) 1995-12-07 1999-08-17 Diversa Corporation Production of enzymes having desired activities by mutagenesis
ATE293684T1 (de) * 1996-01-10 2005-05-15 Novozymes As Methode zur invivo produktion einer mutanten- bibliothek in zellen
US5942430A (en) 1996-02-16 1999-08-24 Diversa Corporation Esterases
US5958751A (en) 1996-03-08 1999-09-28 Diversa Corporation α-galactosidase
US6096548A (en) 1996-03-25 2000-08-01 Maxygen, Inc. Method for directing evolution of a virus
US5783431A (en) * 1996-04-24 1998-07-21 Chromaxome Corporation Methods for generating and screening novel metabolic pathways
US5789228A (en) 1996-05-22 1998-08-04 Diversa Corporation Endoglucanases
US5877001A (en) 1996-06-17 1999-03-02 Diverso Corporation Amidase
US5763239A (en) 1996-06-18 1998-06-09 Diversa Corporation Production and use of normalized DNA libraries
US5939300A (en) 1996-07-03 1999-08-17 Diversa Corporation Catalases
US6093873A (en) 1996-08-19 2000-07-25 Institut National De La Sante Et De La Recherche Medicale Genetically engineered mice containing alterations in the gene encoding RXR
US5834458A (en) 1996-10-09 1998-11-10 Eli Lilly And Company Heterocyclic compounds and their use
CN1208456C (zh) * 1996-12-20 2005-06-29 诺维信公司 体内重组
IL130635A0 (en) * 1997-01-17 2000-06-01 Maxygen Inc Evolution of whole cells and organisms by recursive sequence recombination
US6326204B1 (en) 1997-01-17 2001-12-04 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
DE69839668D1 (de) * 1997-03-18 2008-08-14 Novozymes As Ein in-vitro-verfahren zur herstellung einer dna-bibliothek
IL131986A0 (en) * 1997-03-18 2001-03-19 Novo Nordisk As Shuffling of heterologous dna sequences
AU6611498A (en) * 1997-03-18 1998-10-12 Novo Nordisk A/S Method for constructing a library using dna shuffling
US5948653A (en) 1997-03-21 1999-09-07 Pati; Sushma Sequence alterations using homologous recombination
US6153410A (en) * 1997-03-25 2000-11-28 California Institute Of Technology Recombination of polynucleotide sequences using random or defined primers
US6051049A (en) 1997-05-29 2000-04-18 Exothermic Distribution Corporation Utilization of strontium aluminate in steelmaking
US5948666A (en) 1997-08-06 1999-09-07 Diversa Corporation Isolation and identification of polymerases
US5876997A (en) 1997-08-13 1999-03-02 Diversa Corporation Phytase
JP3712255B2 (ja) 1997-12-08 2005-11-02 カリフォルニア・インスティチュート・オブ・テクノロジー ポリヌクレオチドおよびポリペプチド配列を生成するための方法
US6087341A (en) 1998-02-12 2000-07-11 The Board Of Trustees Of The Leland Standford Junior University Introduction of nucleic acid into skin cells by topical application
US6365408B1 (en) 1998-06-19 2002-04-02 Maxygen, Inc. Methods of evolving a polynucleotides by mutagenesis and recombination
KR20010083086A (ko) 1998-07-28 2001-08-31 추후제출 기능적 진핵세포 단백질의 발현
CA2333914A1 (en) 1998-08-12 2000-02-24 Maxygen, Inc. Dna shuffling to produce herbicide selective crops
JP2002537758A (ja) 1998-09-29 2002-11-12 マキシジェン， インコーポレイテッド コドン変更された遺伝子のシャッフリング
US6376246B1 (en) * 1999-02-05 2002-04-23 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US6397053B1 (en) * 1999-12-30 2002-05-28 Koninklijke Philips Electronics N.V. (Kpenv) Reduction of power consumption with increased standby time in wireless communications device
US20050084907A1 (en) * 2002-03-01 2005-04-21 Maxygen, Inc. Methods, systems, and software for identifying functional biomolecules
US20090312196A1 (en) * 2008-06-13 2009-12-17 Codexis, Inc. Method of synthesizing polynucleotide variants

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101258244B (zh) * 2005-08-08 2012-02-15 基因技术股份公司 用于在体内进行蛋白质的选择性进化的方法
US10202608B2 (en) 2006-08-31 2019-02-12 Gen9, Inc. Iterative nucleic acid assembly using activation of vector-encoded traits
US10457935B2 (en) 2010-11-12 2019-10-29 Gen9, Inc. Protein arrays and methods of using and making the same
US11845054B2 (en) 2010-11-12 2023-12-19 Gen9, Inc. Methods and devices for nucleic acids synthesis
US11084014B2 (en) 2010-11-12 2021-08-10 Gen9, Inc. Methods and devices for nucleic acids synthesis
US10982208B2 (en) 2010-11-12 2021-04-20 Gen9, Inc. Protein arrays and methods of using and making the same
US11702662B2 (en) 2011-08-26 2023-07-18 Gen9, Inc. Compositions and methods for high fidelity assembly of nucleic acids
US10308931B2 (en) 2012-03-21 2019-06-04 Gen9, Inc. Methods for screening proteins using DNA encoded chemical libraries as templates for enzyme catalysis
US10081807B2 (en) 2012-04-24 2018-09-25 Gen9, Inc. Methods for sorting nucleic acids and multiplexed preparative in vitro cloning
US10927369B2 (en) 2012-04-24 2021-02-23 Gen9, Inc. Methods for sorting nucleic acids and multiplexed preparative in vitro cloning
US11072789B2 (en) 2012-06-25 2021-07-27 Gen9, Inc. Methods for nucleic acid assembly and high throughput sequencing
US10662424B2 (en) 2012-12-06 2020-05-26 Agilent Technologies, Inc. Molecular fabrication
US11549106B2 (en) 2012-12-06 2023-01-10 Agilent Technologies, Inc. Molecular fabrication
CN104837994A (zh) * 2012-12-06 2015-08-12 安捷伦科技有限公司 分子制造
CN105934541B (zh) * 2013-11-27 2019-07-12 Gen9股份有限公司 核酸文库及其制造方法
CN105934541A (zh) * 2013-11-27 2016-09-07 Gen9股份有限公司 核酸文库及其制造方法
WO2016058127A1 (zh) * 2014-10-14 2016-04-21 深圳华大基因科技有限公司 一种转座酶打断核酸并加接头的方法和试剂
CN104805508B (zh) * 2015-04-29 2017-07-14 江南大学 一种基于合成单链dna文库进化代谢途径的方法
CN104805508A (zh) * 2015-04-29 2015-07-29 江南大学 一种基于合成单链dna文库进化代谢途径的方法
CN110366593A (zh) * 2017-02-21 2019-10-22 苏黎世联邦理工学院 Dna零件，途径和基因组的进化指导的多重dna装配

Also Published As

Publication number Publication date
US20080261833A1 (en) 2008-10-23
DE69525084D1 (de) 2002-02-28
DE69525084T2 (de) 2002-07-11
US5605793A (en) 1997-02-25
JP2003033180A (ja) 2003-02-04
DK0752008T3 (da) 2002-06-17
DE69526497T2 (de) 2002-11-21
JP2003174881A (ja) 2003-06-24
US5811238A (en) 1998-09-22
EP0934999A1 (en) 1999-08-11
US6602986B1 (en) 2003-08-05
EP1094108A3 (en) 2001-05-09
EP0752008B2 (en) 2013-04-10
DE752008T1 (de) 2001-04-05
CA2182393C (en) 2005-05-17
US6297053B1 (en) 2001-10-02
US6277638B1 (en) 2001-08-21
EP1205547A2 (en) 2002-05-15
WO1995022625A1 (en) 1995-08-24
US20040241853A1 (en) 2004-12-02
JPH10500561A (ja) 1998-01-20
EP0752008A1 (en) 1997-01-08
AU703264B2 (en) 1999-03-25
JP2001057893A (ja) 2001-03-06
EP1205547A3 (en) 2002-07-24
US7981614B2 (en) 2011-07-19
AU703264C (en) 2003-06-12
CA2182393A1 (en) 1995-08-24
US6287861B1 (en) 2001-09-11
DE69526497D1 (de) 2002-05-29
US7288375B2 (en) 2007-10-30
ATE211761T1 (de) 2002-01-15
JP2004254709A (ja) 2004-09-16
EP0934999B1 (en) 2002-01-09
DE934999T1 (de) 2001-04-05
EP1094108A2 (en) 2001-04-25
DE1094108T1 (de) 2002-10-17
AU2010200270A1 (en) 2010-02-18
AU2971495A (en) 1995-09-04
RU2157851C2 (ru) 2000-10-20
EP0752008B1 (en) 2002-04-24
US5830721A (en) 1998-11-03
DK0934999T3 (da) 2002-02-18
US6444468B1 (en) 2002-09-03
JP2007244399A (ja) 2007-09-27
ES2165652T3 (es) 2002-03-16
JP3485560B2 (ja) 2004-01-13
JP3393848B2 (ja) 2003-04-07
US6576467B1 (en) 2003-06-10
US6420175B1 (en) 2002-07-16
AU2006202542A1 (en) 2006-07-13
US20100222237A1 (en) 2010-09-02
US6132970A (en) 2000-10-17
EP0752008A4 (en) 1999-03-17
ATE216722T1 (de) 2002-05-15
JP4067516B2 (ja) 2008-03-26
ES2176324T3 (es) 2002-12-01

Similar Documents

Publication Publication Date Title
CN1145641A (zh) 通过随机片段化和重装配进行的dna诱变
Patten et al. Applications of DNA shuffling to pharmaceuticals and vaccines
CN1484698A (zh) 具有c型类凝集素结构域的支架结构的蛋白质组合文库
CN111757937A (zh) 腺苷碱基编辑器的用途
CN1230990A (zh) 用随机或确定的引物来重组多核苷酸序列
CN1757724A (zh) 具有独特特异性的多个重组位点在重组克隆中的用途
CN1173878A (zh) 双功能或双价抗体片段类似物
CN1599751A (zh) 分枝杆菌比较基因组学作为鉴定分枝杆菌病的诊断、预防或治疗靶的工具
CN1836041A (zh) 浏览突变
CN87105787A (zh) 纯化的热稳定性酶以及用其进行扩增、检测和/或克隆核酸顺序的方法
JPH05503000A (ja) 新規の遺伝子及びポリペチドの無細胞合成並びに単離
CN1956996A (zh) 基于泛素蛋白的人工结合蛋白的生产
CN85109740A (zh) 能表达脱辅基水母发光蛋白(Apoaeq-uorin)的重组DNA载体
McClain Seven terminal steps in a biosynthetic pathway leading from DNA to transfer RNA
CN1529759A (zh) 模块转染系统
CN1264860C (zh) 在遗传载体上表面展示蛋白的方法
CN1449444A (zh) 无细胞蛋白质合成用转录模型的设计、结构及使用其合成无细胞麦胚芽蛋白质的稀释批式法
CN1590407A (zh) 多功能性造血受体激动剂
Oosting Molecular biotechnology: from dna sequence to therapeutic protein
EP1356090A2 (en) Sequence based screening
CN1649626A (zh) 可与触珠蛋白受体配体结合相互作用的分子的用途
CN1289668C (zh) 一种用于抗体改形的体外分子定向进化方法
EP1670932B1 (en) Libraries of recombinant chimeric proteins
CN1894272A (zh) 蛋白质表达
CN1498276A (zh) 通过三螺旋相互作用纯化与检测双链dna靶序列的方法

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication