WO2002000879A3 - Methods for producing modified glycoproteins - Google Patents

Methods for producing modified glycoproteins Download PDF

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Publication number
WO2002000879A3
WO2002000879A3 PCT/US2001/020553 US0120553W WO0200879A3 WO 2002000879 A3 WO2002000879 A3 WO 2002000879A3 US 0120553 W US0120553 W US 0120553W WO 0200879 A3 WO0200879 A3 WO 0200879A3
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WO
WIPO (PCT)
Prior art keywords
glycoproteins
human
produce
glycans
fungi
Prior art date
Application number
PCT/US2001/020553
Other languages
French (fr)
Other versions
WO2002000879A2 (en
Inventor
Tillman U Gerngross
Original Assignee
Glycofi Inc
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=27395978&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=WO2002000879(A3) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority to MXPA03000105A priority Critical patent/MXPA03000105A/en
Priority to NZ523476A priority patent/NZ523476A/en
Priority to AT01954606T priority patent/ATE309385T1/en
Priority to KR1020027017911A priority patent/KR100787073B1/en
Priority to CA002412701A priority patent/CA2412701A1/en
Application filed by Glycofi Inc filed Critical Glycofi Inc
Priority to AU2001276842A priority patent/AU2001276842B2/en
Priority to AU7684201A priority patent/AU7684201A/en
Priority to JP2002506194A priority patent/JP2004501642A/en
Priority to EP01954606A priority patent/EP1297172B1/en
Priority to DE60114830T priority patent/DE60114830T2/en
Priority to DK04025648T priority patent/DK1522590T3/en
Publication of WO2002000879A2 publication Critical patent/WO2002000879A2/en
Publication of WO2002000879A3 publication Critical patent/WO2002000879A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/005—Glycopeptides, glycoproteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • 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)
    • C12N9/2477—Hemicellulases not provided in a preceding group
    • C12N9/2488—Mannanases
    • 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)
    • C12Y302/01113—Mannosyl-oligosaccharide 1,2-alpha-mannosidase (3.2.1.113), i.e. alpha-1,2-mannosidase
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/04—Fusion polypeptide containing a localisation/targetting motif containing an ER retention signal such as a C-terminal HDEL motif
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/05—Fusion polypeptide containing a localisation/targetting motif containing a GOLGI retention signal
    • 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)

Abstract

Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce 'human-like' glycoproteins.
PCT/US2001/020553 2000-06-28 2001-06-27 Methods for producing modified glycoproteins WO2002000879A2 (en)

Priority Applications (11)

Application Number Priority Date Filing Date Title
DK04025648T DK1522590T3 (en) 2000-06-28 2001-06-27 Process for Preparation of Modified Glycoproteins
AU7684201A AU7684201A (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
AT01954606T ATE309385T1 (en) 2000-06-28 2001-06-27 METHOD FOR PRODUCING MODIFIED GLYCOPROTEINS
KR1020027017911A KR100787073B1 (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
CA002412701A CA2412701A1 (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
MXPA03000105A MXPA03000105A (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins.
AU2001276842A AU2001276842B2 (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
NZ523476A NZ523476A (en) 2000-06-28 2001-06-27 Methods for humanizing glycosylation of recombinant glycoproteins expressed in lower eukaryotes
JP2002506194A JP2004501642A (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
EP01954606A EP1297172B1 (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins
DE60114830T DE60114830T2 (en) 2000-06-28 2001-06-27 PROCESS FOR PRODUCING MODIFIED GLYCOPROTEINS

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US21435800P 2000-06-28 2000-06-28
US60/214,358 2000-06-28
US21563800P 2000-06-30 2000-06-30
US60/215,638 2000-06-30
US27999701P 2001-03-30 2001-03-30
US60/279,997 2001-03-30

Publications (2)

Publication Number Publication Date
WO2002000879A2 WO2002000879A2 (en) 2002-01-03
WO2002000879A3 true WO2002000879A3 (en) 2002-09-06

Family

ID=27395978

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2001/020553 WO2002000879A2 (en) 2000-06-28 2001-06-27 Methods for producing modified glycoproteins

Country Status (15)

Country Link
US (13) US7029872B2 (en)
EP (5) EP2322644A1 (en)
JP (2) JP2004501642A (en)
KR (1) KR100787073B1 (en)
AT (2) ATE440959T1 (en)
AU (2) AU2001276842B2 (en)
CA (1) CA2412701A1 (en)
CY (1) CY1109639T1 (en)
DE (2) DE60114830T2 (en)
DK (2) DK1297172T3 (en)
ES (2) ES2252261T3 (en)
MX (1) MXPA03000105A (en)
NZ (1) NZ523476A (en)
PT (1) PT1522590E (en)
WO (1) WO2002000879A2 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8877462B2 (en) 2000-06-28 2014-11-04 Glycofi, Inc. Combinatorial DNA library for producing modified N-glycans in lower eukaryotes
US8986949B2 (en) 2003-02-20 2015-03-24 Glycofi, Inc. Endomannosidases in the modification of glycoproteins in eukaryotes
US9574218B2 (en) 2001-01-19 2017-02-21 Phyton Holdings, Llc Method of co-expressing galactosyltransferase and a glycoprotein in a transgenic plant cell and sialylating the glycoprotein for production of glycoprotein having human-type sugar chain

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