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Publication numberUS20070244047 A1
Publication typeApplication
Application numberUS 11/714,841
Publication dateOct 18, 2007
Filing dateMar 7, 2007
Priority dateDec 21, 2001
Also published asCA2471363A1, CA2471363C, CA2841097A1, EP1463751A2, EP1463751A4, EP1463751B1, EP1997829A1, EP2261250A1, EP2277888A2, EP2277888A3, EP2277889A2, EP2277889A3, EP2277910A1, US7141547, US7238667, US7592010, US7799759, US7847079, US8012464, US8071539, US8211439, US8252739, US8513189, US20050186664, US20060194735, US20060276396, US20070259815, US20080146503, US20080153751, US20080161243, US20080167239, US20080167240, US20080213886, US20090093402, US20090099073, US20110009312, US20120046221, US20130150296, US20140179596, WO2003060071A2, WO2003060071A3
Publication number11714841, 714841, US 2007/0244047 A1, US 2007/244047 A1, US 20070244047 A1, US 20070244047A1, US 2007244047 A1, US 2007244047A1, US-A1-20070244047, US-A1-2007244047, US2007/0244047A1, US2007/244047A1, US20070244047 A1, US20070244047A1, US2007244047 A1, US2007244047A1
InventorsCraig Rosen, William Haseltine, Steven Ruben
Original AssigneeHuman Genome Sciences, Inc., Delta Biotechnology Limited
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Albumin fusion proteins
US 20070244047 A1
Abstract
The present invention encompasses albumin fusion proteins. Nucleic acid molecules encoding the albumin fusion proteins of the invention are also encompassed by the invention, as are vectors containing these nucleic acids, host cells transformed with these nucleic acids vectors, and methods of making the albumin fusion proteins of the invention and using these nucleic acids, vectors, and/or host cells. Additionally the present invention encompasses pharmaceutical compositions comprising albumin fusion proteins and methods of treating, preventing, or ameliorating diseases, disorders or conditions using albumin fusion proteins of the invention.
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Claims(20)
1. An albumin fusion protein comprising an exendin-4 polypeptide fused to an albumin polypeptide, wherein said exendin-4 polypeptide is selected from:
(a) a full-length exendin-4;
(b) a mature exendin-4;
(c) a fragment of exendin-4, wherein said fragment has exendin-4 activity;
(d) a polypeptide consisting of an amino acid sequence at least 90% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(e) a polypeptide consisting of an amino acid sequence at least 95% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(f) a polypeptide consisting of an amino acid sequence at least 99% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(g) a polypeptide comprising amino acids 1 to 87 of SEQ ID NO:635;
(h) a polypeptide comprising amino acids 48 to 86 of SEQ ID NO:635; and wherein said albumin polypeptide is selected from:
(i) a full-length albumin;
(ii) a mature albumin;
(iii) a fragment of albumin, wherein said fragment of albumin has the ability to prolong the serum half-life of the exendin-4 polypeptide;
(iv) a polypeptide consisting of an amino acid sequence at least 95% identical to albumin, wherein said polypeptide has the ability to prolong the serum half-life of the exendin-4 polypeptide;
(v) amino acids 1 to 585 of SEQ ID NO:1038; and
(vi) amino acids 1 to 387 of SEQ ID NO: 1038.
2. The albumin fusion protein of claim 1, produced from a host cell comprising an albumin fusion construct which expresses said albumin fusion protein, wherein said albumin fusion construct is selected from:
(a) 2469; and
(b) 2470.
3. The albumin fusion protein of claim 1, wherein said albumin fusion protein comprises an amino acid sequence selected from:
(a) amino acids 25 to 648 of SEQ ID NO:419; and
(b) amino acids 25 to 648 of SEQ ID NO:420.
4. The albumin fusion protein of claim 1, wherein said exendin-4 polypeptide is fused to the N-terminus, the C-terminus, or both the N-terminus and C-terminus of said albumin polypeptide.
5. The albumin fusion protein of claim 1, which is expressed in a host cell.
6. The albumin fusion protein of claim 5, wherein said host cell is a yeast, mammalian, or bacterial cell.
7. A nucleic acid molecule comprising a first polynucleotide and a second polynucleotide, wherein said first polynucleotide encodes an exendin-4 polypeptide selected from:
(a) a full-length exendin-4;
(b) a mature exendin-4;
(c) a fragment of exendin-4, wherein said fragment has exendin-4 activity;
(d) a polypeptide consisting of an amino acid sequence at least 90% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(e) a polypeptide consisting of an amino acid sequence at least 95% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(f) a polypeptide consisting of an amino acid sequence at least 99% identical to exendin-4, wherein said polypeptide has exendin-4 activity;
(g) a polypeptide comprising amino acids 1 to 87 of SEQ ID NO:635;
(h) a polypeptide comprising amino acids 48 to 86 of SEQ ID NO:635; and wherein said second polynucleotide encodes an albumin polypeptide selected from:
(i) a full-length albumin;
(ii) a mature albumin;
(iii) a fragment of albumin, wherein said fragment of albumin has the ability to prolong the serum half-life of the exendin-4 polypeptide;
(iv) a polypeptide consisting of an amino acid sequence at least 95% identical to albumin, wherein said polypeptide has the ability to prolong the serum half-life of the exendin-4 polypeptide;
(v) amino acids 1 to 585 of SEQ ID NO:1038; and
(vi) amino acids 1 to 387 of SEQ ID NO:1038.
8. An albumin fusion construct comprising the nucleic acid molecule of claim 7, which upon expression in a host cell produces an albumin fusion protein comprising exendin-4 fused to human albumin or a human albumin fragment or variant, wherein said albumin fusion construct is selected from:
(a) 2469; and
(b) 2470.
9. A vector comprising the nucleic acid molecule of claim 7.
10. An isolated host cell comprising the nucleic acid molecule of claim 7.
11. An isolated host cell comprising the vector of claim 9.
12. An isolated host cell comprising the albumin fusion construct of claim 8.
13. A method for producing a fusion protein, comprising:
(a) culturing the host cell of claim 10 under conditions suitable to produce the fusion protein encoded by said polynucleotide; and
(b) recovering said fusion protein.
14. A method for producing a fusion protein, comprising:
(a) culturing the host cell of claim 11 under conditions suitable to produce the fusion protein encoded by said vector; and
(b) recovering said fusion protein.
15. A method for producing a fusion protein, comprising:
(a) culturing the host cell of claim 12 under conditions suitable to produce the fusion protein encoded by said albumin fusion construct; and
(b) recovering said fusion protein.
16. A method of treating a disease or disorder in a patient comprising the step of administering an effective amount of the albumin fusion protein of claim 1.
17. The method of claim 16, wherein the disease or disorder is selected from: hyperglycemia, diabetes, diabetes insipidus, diabetes mellitus, type 1 diabetes, type 2 diabetes, insulin resistance, insulin deficiency, hyperlipidemia, hyperketonemia, non-insulin dependent diabetes mellitus (NIDDM), insulin-dependent diabetes mellitus (IDDM), obesity, heart disease, infections, retinopathy, ulcers, metabolic disorders, immune disorders, vascular disorders, suppression of body weight, suppression of appetite, and syndrome X.
18. The method of claim 17, wherein the disease or disorder is diabetes.
19. The method of claim 17, wherein the disease or disorder is obesity or of losing weight in a patient.
20. A method of stimulating insulin synthesis and release, enhancing adipose, muscle or liver tissue sensitivity towards insulin uptake, stimulating glucose uptake, slowing digestive process, or blocking the secretion of glucagon in a patient, comprising administering to said patient an albumin fusion protein of claim 1.
Description
CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional of U.S. application Ser. No. 11/429,276, filed May 8, 2006, which is a continuation of U.S. application Ser. No. 10/775,204, filed Feb. 11, 2004, which is a continuation of International Application No. PCT/US02/40891, filed Dec. 23, 2002, which claims benefit under 35 USC 119(e) of U.S. Provisional Application Nos. 60/341,811, filed Dec. 21, 2001; 60/350,358, filed Jan. 24, 2002; 60/351,360, filed Jan. 28, 2002; 60/359,370, filed Feb. 26, 2002; 60/360,000, filed Feb. 28, 2002; 60/367,500, filed Mar. 27, 2002; 60/370,227, filed Apr. 8, 2002; 60/378,950, filed May 10, 2002; 60/382,617, filed May 24, 2002; 60/383,123, filed May 28, 2002; 60/385,708, filed Jun. 5, 2002; 60/394,625, filed Jul. 10, 2002; 60/398,008, filed Jul. 24, 2002; 60/402,131, filed Aug. 9, 2002; 60/402,708, filed Aug. 13, 2002; 60/411,426, filed Sep. 18, 2002; 60/411,355, filed Sep. 18, 2002; 60/414,984, filed Oct. 2, 2002; 60/417,611, filed Oct. 11, 2002; 60/420,246, filed Oct. 23, 2002; and 60/423,623, filed Nov. 5, 2002. All of the above listed applications are incorporated by reference herein.

REFERENCE TO SEQUENCE LISTING ON COMPACT DISC

This application refers to a “Sequence Listing” listed below, which is provided as an electronic document on three identical compact disc (CD-R), labeled “Copy 1,” “Copy 2,” and “CRF.” These compact discs each contain the file “PF564C2D1 SEQLIST FINAL.txt” (3,568,877 bytes, created on Mar. 6, 2007), which is incorporated by reference in its entirety. The Sequence Listing may be viewed on an IBM-PC machine running the MS-Windows operating system.

BACKGROUND OF THE INVENTION

The invention relates generally to Therapeutic proteins (including, but not limited to, at least one polypeptide, antibody, peptide, or fragment and variant thereof) fused to albumin or fragments or variants of albumin. The invention encompasses polynucleotides encoding therapeutic albumin fusion proteins, therapeutic albumin fusion proteins, compositions, pharmaceutical compositions, formulations and kits. Host cells transformed with the polynucleotides encoding therapeutic albumin fusion proteins are also encompassed by the invention, as are methods of making the albumin fusion proteins of the invention using these polynucleotides, and/or host cells.

Human serum albumin (HSA, or HA), a protein of 585 amino acids in its mature form (as shown in FIG. 1 (SEQ ID NO:1038)), is responsible for a significant proportion of the osmotic pressure of serum and also functions as a carrier of endogenous and exogenous ligands. At present, HA for clinical use is produced by extraction from human blood. The production of recombinant HA (rHA) in microorganisms has been disclosed in EP 330 451 and EP 361 991.

Therapeutic proteins in their native state or when recombinantly produced, such as interferons and growth hormones, are typically labile molecules exhibiting short shelf-lives, particularly when formulated in aqueous solutions. The instability in these molecules when formulated for administration dictates that many of the molecules must be lyophilized and refrigerated at all times during storage, thereby rendering the molecules difficult to transport and/or store. Storage problems are particularly acute when pharmaceutical formulations must be stored and dispensed outside of the hospital environment.

Few practical solutions to the storage problems of labile protein molecules have been proposed. Accordingly, there is a need for stabilized, long lasting formulations of proteinaceous therapeutic molecules that are easily dispensed, preferably with a simple formulation requiring minimal post-storage manipulation.

SUMMARY OF THE INVENTION

The present invention encompasses albumin fusion proteins comprising a Therapeutic protein (e.g., a polypeptide, antibody, or peptide or fragment or variant thereof) fused to albumin or a fragment (portion) or variant of albumin. The present invention also encompasses polynucleotides comprising, or alternatively consisting of, nucleic acid molecules encoding a Therapeutic protein (e.g., a polypeptide, antibody; or peptide, or fragment or variant thereof) fused to albumin or a fragment (portion) or variant of albumin. The present invention also encompasses polynucleotides, comprising, or alternatively consisting of; nucleic acid molecules encoding proteins comprising a Therapeutic protein (e.g., a polypeptide, antibody, or peptide, or fragment or variant thereof) fused to albumin or a fragment (portion) or variant of albumin, that is sufficient to prolong the shelf life of the Therapeutic protein, and/or stabilize the Therapeutic protein and/or its activity in solution (or in a pharmaceutical composition) in vitro and/or in vivo. Albumin fusion proteins encoded by a polynucleotide of the invention are also encompassed by the invention, as are host cells transformed with polynucleotides of the invention, and methods of making the albumin fusion proteins of the invention and using these polynucleotides of the invention, and/or host cells.

In a preferred aspect of the invention, albumin fusion proteins include, but are not limited to, those encoded by the polynucleotides described in Table 2.

The invention also encompasses pharmaceutical formulations comprising an albumin fusion protein of the invention and a pharmaceutically acceptable diluent or carrier. Such formulations may be in a kit or container. Such kit or container may be packaged with instructions pertaining to the extended shelf life of the Therapeutic protein. Such formulations may be used in methods of treating, preventing, ameliorating or diagnosing a disease or disease symptom in a patient, preferably a mammal, most preferably a human, comprising the step of administering the pharmaceutical formulation to the patient.

In other embodiments, the present invention encompasses methods of preventing, treating, or ameliorating a disease or disorder. In preferred embodiments, the present invention encompasses a method of treating a disease or disorder listed in the “Preferred Indication: Y” column of Table 1 comprising administering to a patient in which such treatment, prevention or amelioration is desired an albumin fusion protein of the invention that comprises a Therapeutic protein or portion corresponding to a Therapeutic protein (or fragment or variant thereof) disclosed in the “Therapeutic Protein: X” column of Table 1 (in the same row as the disease or disorder to be treated is listed in the “Preferred Indication: Y” column of Table 1) in an amount effective to treat, prevent or ameliorate the disease or disorder.

In one embodiment, an albumin fusion protein described in Table 1 or 2 has extended shelf life.

In a second embodiment, an albumin fusion protein described in Table 1 or 2 is more stable than the corresponding unfused Therapeutic molecule described in Table 1.

The present invention further includes transgenic organisms modified to contain the nucleic acid molecules of the invention (including, but not limited to, the polynucleotides described in Tables 1 and 2), preferably modified to express an albumin fusion protein of the invention.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A-D shows the amino acid sequence of the mature form of human albumin (SEQ ID NO:1038) and a polynucleotide encoding it (SEQ ID NO:1037).

FIG. 2 shows the restriction map of the pPPC0005 cloning vector ATCC deposit PTA-3278.

FIG. 3 shows the restriction map of the pSAC35 yeast S. cerevisiae expression vector (Sleep et al., BioTechnology 8:42 (1990)).

FIG. 4 shows the effect of various dilutions of EPO albumin fusion proteins encoded by DNA comprised in Construct ID NOS. (hereinafter CID) 1966 and 1981 and recombinant human EPO on the proliferation of TF-1 cells (see Examples 8 and 9). Cells were washed 3× to remove GM-CSF and plated at 10,000 cells/well for 72 hours in the presence of 3-fold dilutions of CID 1966 protein or CID 1981 protein. Concentrations used were calculated based on the weight of Epo alone, not HSA plus Epo. Recombinant human Epo (rhEpo) was used as the positive control and serially diluted 3 fold from 100 ng/ml to 0.01 ng/ml. Cells were exposed to 0.5 mCi/well of 3H-thymidine for an additional 18 hours. (□) rhEpo; (▾) HSA-Epo 1981; (•) Epo-HSA 1966.

FIG. 5 is a dose response analysis and shows the effect of various doses of recombinant human EPO and EPO albumin fusion proteins encoded by DNA comprised in CID 1966 and 1981 on the percent change in hematocrit from day 0 to day 7 (see Examples 8 and 9). 48 eight-week old female DBA/2NHsd mice were divided into 12 groups of 4 animals each. Recombinant human Epo (rhEpo) was administered subcutaneously at 0.5, 1.5, 4.5 and 12 μg/kg on days 0, 2, 4, and 6. Epo albumin fusion proteins made from constructs CID 1966 and CID 1981 were administered subcutaneously at 2, 6, 18, and 54 μg/kg on days 0, 2, 4, and 6. The higher doses of the Epo albumin fusion proteins-allows a rough equimolar comparison with recombinant human Epo (note that the weight of the fusions is about 3.35 times the weight of non-glycosylated Epo). On days 0 and 7 of the experiment, the animals were bled via a tail vein and the hematocrit was determined by centrifugation. (▪) rhEpo; (∘) CID 1981; (▴) CID 1966.

FIG. 6A shows the effect of various subcutaneous administrations of Epo albumin fusion proteins encoded by DNA comprised in CID 1966 and 1997, respectively, on the percent change in hematocrit from day 0 to day 8 (see Examples 8 and 10). *, p<0.005 compared to rhEpo as determined by Mann-Whitney nonparametric analysis (n=6).

FIG. 6B shows the effect of subcutaneous administrations of Epo albumin fusion proteins encoded by DNA comprised in CID 1997 and 1966 on the percent change in hematocrit from day 0 to day 14 (see Examples 8 and 10). *, p<0.005 compared to rhEpo as determined by Mann-Whitney nonparametric analysis (n=6); **, p<0.05 compared to rhEpo as determined by Mann-Whitney nonparametric analysis (n=6).

FIG. 7 shows the effect of various dilutions albumin fusion proteins encoded by DNA comprised in CID 1981 and 1997, respectively, on the proliferation of TF-1 cells (see Examples 9 and 10). Cells were washed 3× to remove GM-CSF and plated at 10,000 cells/well for 72 hours in the presence of 3-fold dilutions of Epo albumin fusion proteins encoded by CID 1981 or 1997. Equimolar amounts of rhEpo were used as a positive control (4.35 times less protein added since weight of non-glycosylated Epo is 20 kd, while Epo albumin fusion proteins are 87 kd). Cells were exposed to 0.5 μCi/well of 3H-thymidine for an additional 24 hours. (▪) rhEpo Standard; (▴) CID 1981 (CHO); (∘) CID 1997 (NSO).

FIG. 8 shows the effect of various doses of recombinant human EPO (rhEpo) and EPO albumin fusion protein encoded by DNA comprised in construct 1997 (CID 1997) on the percent change in hematocrit from day 0 to day 8 (see Example 10). (▴)=rhEpo, ( )=CID 1997.

FIG. 9 shows the effect of various dilutions of IL2 albumin fusion proteins encoded by DNA comprised in CID 1812 (see Example 15) on CTLL-2 proliferation. 1×104 cells/well were seeded in a 96-well plate in a final volume of 200 ul of complete medium containing the indicated amount of IL2 albumin fusion protein (CID 1812). All samples were run in triplicate. The cells were incubated for 40 hours at 37° C., then 20 ul of Alamar Blue was added and cells incubated for 8 hours. Absorbance at 530/590 was used as a measure of proliferation. EC50=0.386±0.021. (Δ)=CID 1812.

FIG. 10 shows the effect of IL2 albumin fusion protein encoded by DNA comprised in CID 1812 on RENCA tumor growth at day 21 (see Example 15). BALB/c mice (n=10) were injected SC (midflank) with 105 RENCA cells. 10 days later mice received 2 cycles (Day 10 to Day 14 and Days 17-21) of daily (QD) injections of rlL2 (0.9 mg/kg), IL2 albumin fusion protein (CID 1812 protein: 0.6 mg/kg) or PBS (Placebo) or injections every other day (QOD) of CID 1812 protein (0.6 mg/kg). The tumor volume was determined on Day 21 after RENCA inoculation. The data are presented in scatter analysis (each dot representing single animal). Mean value of each group is depicted by horizontal line. *, p=0.0035 between placebo control and CID 1812 protein. The number in parentheses indicates number of mice alive over the total number of mice per group. (◯)=Placebo; (●)=IL2; (Δ)=CID 1812 protein (QD); (□)=CID 1812 protein (QOD).

FIG. 11 shows the effect of various dilutions of GCSF albumin fusion proteins encoded by DNA comprised in CID 1642 and 1643 on NFS-60 cell proliferation (see Examples 19 and 20). (▪)=CID 1642; (▴)=CID 1643; (◯)=HSA.

FIG. 12 shows the effect of recombinant human GCSF (Neupogen) and GCSF albumin fusion protein on total white blood cell count (see Example 19). Total WBC (103 cells/ul) on each day are presented as the group mean ±SEM. GCSF albumin fusion protein was administered sc at either 25 or 100 ug/kg every 4 days×4 (Q4D), or at 100 ug/kg every 7 days×2 (Q7D). Data from Days 8 and 9 for GCSF albumin fusion protein 100 ug/kg Q7 are presented as Days 9 and 10, respectively, to facilitate comparison with other groups. Controls were saline vehicle administered SC every 4 days×4 (Vehicle Q4D), or Neupogen administered SC daily×14 (Neupogen 5 ug/kg QD). The treatment period is considered Days 1-14, and the recovery period, Days 15-28.

FIG. 13 shows the effect of various dilutions of IFNb albumin fusion proteins encoded by DNA comprised in CID 2011 and 2053 on SEAP activity in the ISRE-SEAP/293F reporter cells (see Example 25). Proteins were serially diluted from 5e-7 to 1e-14 g/ml in DMEM/10% FBS and used to treat ISRE-SEAP/293F reporter cells. After 24 hours supernatants were removed from reporter cells and assayed for SEAP activity. IFNb albumin fusion protein was purified from three stable clones: 293F/#2011, CHO/#2011 and NSO/#2053. Mammalian derived IFNb, Avonex, came from Biogen and was reported to have a specific activity of 2.0e5 IU/ug.

FIG. 14 illustrates the steady-state levels of insulin mRNA in INS-1 (832/13) cells after treatment with GLP-1 or GLP-1 albumin fusion protein encoded by construct ID 3070 (CID 3070 protein). Both GLP-1 and the CID 3070 protein stimulate transcription of the insulin gene in INS-1 cells. The first bar (black) represents the untreated cells. Bars 2-4 (white) represent cells treated with the indicated concentrations of GLP-1. Bars 5-7 (gray) represent cells treated with the indicated concentrations of CID 3070 protein.

FIG. 15 compares the anti-proliferative activity of IFN albumin fusion protein encoded by CID 3165 (CID 3165 protein) and recombinant IFNa (rIFNa) on Hs294T melanoma cells. The cells were cultured with varying concentrations of either CID 3165 protein or rIFNa and proliferation was measured by BrdU incorporation after 3 days of culture. CID 3165 protein caused measurable inhibition of cell proliferation at concentrations above 10 ng/ml with 50% inhibition achieved at approximately 200 ng/ml. (▪)=CID 3165 protein, (♦)=rIFNa.

FIG. 16 shows the effect of various dilutions of IFNa albumin fusion proteins on SEAP activity in the ISRE-SEAP/293F reporter cells. One preparation of IFNa fused upstream of albumin (♦) was tested, as well as two different preparations of IFNa fused downstream of albumin (▴) and (▪).

FIG. 17 shows the effect of time and dose of IFNa albumin fusion protein encoded by DNA comprised in construct 2249 (CID 2249 protein) on the mRNA level of OAS (p41) in treated monkeys (see Example 31). Per time point: first bar=Vehicle control, 2nd bar=30 ug/kg CID 2249 protein day 1 iv, third bar=30 ug/kg CID 2249 protein day 1 sc, 4th bar=300 ug/kg CID 2249 protein day 1 sc, 5th bar=40 ug/kg recombinant IFNa day 1, 3 and 5 sc.

FIG. 18 shows the effect of various dilutions of insulin albumin fusion proteins encoded by DNA comprised in constructs 2250 and 2276 on glucose uptake in 3T3-L1 adipocytes (see Examples 33 and 35).

FIG. 19 shows the effect of various GCSF albumin fusion proteins, including those encoded by CID #1643 and #2702 (L-171, see Example 114), on NFS cell proliferation. The horizontal dashed line indicates the minimum level of detection.

DETAILED DESCRIPTION

Definitions

The following definitions are provided to facilitate understanding of certain terms used throughout this specification.

As used herein, “polynucleotide” refers to a nucleic acid molecule having a nucleotide sequence encoding a fusion protein comprising, or alternatively consisting of, at least one molecule of albumin (or a fragment or variant thereof) joined in frame to at least one Therapeutic protein X (or fragment or variant thereof); a nucleic acid molecule having a nucleotide sequence encoding a fusion protein comprising, or alternatively consisting of, the amino acid sequence of SEQ ID NO:Y (as described in column 6 of Table 2) or a fragment or variant thereof; a nucleic acid molecule having a nucleotide sequence comprising or alternatively consisting of the sequence shown in SEQ ID NO:X; a nucleic acid molecule having a nucleotide sequence encoding a fusion protein comprising, or alternatively consisting of, the amino acid sequence of SEQ ID NO:Z; a nucleic acid molecule having a nucleotide sequence encoding an albumin fusion protein of the invention generated as described in Table 2 or in the Examples; a nucleic acid molecule having a nucleotide sequence encoding a Therapeutic albumin fusion protein of the invention, a nucleic acid molecule having a nucleotide sequence contained in an albumin fusion construct described in Table 2, or a nucleic acid molecule having a nucleotide sequence contained in an albumin fusion construct deposited with the ATCC (as described in Table 3).

As used herein, “albumin fusion construct” refers to a nucleic acid molecule comprising, or alternatively consisting of, a polynucleotide encoding at least one molecule of albumin (or a fragment or variant thereof) joined in frame to at least one polynucleotide encoding at least one molecule of a Therapeutic protein (or fragment or variant thereof); a nucleic acid molecule comprising, or alternatively consisting of, a polynucleotide encoding at least one molecule of albumin (or a fragment or variant thereof) joined in frame to at least one polynucleotide encoding at least one molecule of a Therapeutic protein (or fragment or variant thereof) generated as described in Table 2 or in the Examples; or a nucleic acid molecule comprising, or alternatively consisting of, a polynucleotide encoding at least one molecule of albumin (or a fragment or variant thereof) joined in frame to at least one polynucleotide encoding at least one molecule of a Therapeutic protein (or fragment or variant thereof), further comprising, for example, one or more of the following elements: (1) a functional self-replicating vector (including but not limited to, a shuttle vector, an expression vector, an integration vector, and/or a replication system), (2) a region for initiation of transcription (e.g., a promoter region, such as for example, a regulatable or inducible promoter, a constitutive promoter), (3) a region for termination of transcription, (4) a leader sequence, and (5) a selectable marker. The polynucleotide encoding the Therapeutic protein and albumin protein, once part of the albumin fusion construct, may each be referred to as a “portion,” “region” or “moiety” of the albumin fusion construct.

The present invention relates generally to polynucleotides encoding albumin fusion proteins; albumin fusion proteins; and methods of treating, preventing, or ameliorating diseases or disorders using albumin fusion proteins or polynucleotides encoding albumin fusion proteins. As used herein, “albumin fusion protein” refers to a protein formed by the fusion of at least one molecule of albumin (or a fragment or variant thereof) to at least one molecule of a Therapeutic protein (or fragment or variant thereof). An albumin fusion protein of the invention comprises at least a fragment or variant of a Therapeutic protein and at least a fragment or variant of human serum albumin, which are associated with one another by genetic fusion (i.e., the albumin fusion protein is generated by translation of a nucleic acid in which a polynucleotide encoding all or a portion of a Therapeutic protein is joined in-frame with a polynucleotide encoding all or a portion of albumin). The Therapeutic protein and albumin protein, once part of the albumin fusion protein, may each be referred to as a “portion”, “region” or “moiety” of the albumin fusion protein (e.g., a “Therapeutic protein portion” or an “albumin protein portion”). In a highly preferred embodiment, an albumin fusion protein of the invention comprises at least one molecule of a Therapeutic protein X or fragment or variant of thereof (including, but not limited to a mature form of the Therapeutic protein X) and at least one molecule of albumin or fragment or variant thereof (including but not limited to a mature form of albumin).

In a further preferred embodiment, an albumin fusion protein of the invention is processed by a host cell and secreted into the surrounding culture medium. Processing of the nascent albumin fusion protein that occurs in the secretory pathways of the host used for expression may include, but is not limited to signal peptide cleavage; formation of disulfide bonds; proper folding; addition and processing of carbohydrates (such as for example, N- and O-linked glycosylation); specific proteolytic cleavages; and assembly into multimeric proteins. An albumin fusion protein of the invention is preferably in the processed form. In a most preferred embodiment, the “processed form of an albumin fusion protein” refers to an albumin fusion protein product which has undergone N-terminal signal peptide cleavage, herein also referred to as a “mature albumin fusion protein”.

In several instances, a representative clone containing an albumin fusion construct of the invention was deposited with the American Type Culture Collection (herein referred to as “ATCC®”). Furthermore, it is possible to retrieve a given albumin fusion construct from the deposit by techniques known in the art and described elsewhere herein. The ATCC® is located at 10801 University Boulevard, Manassas, Va. 20110-2209, USA. The ATCC® deposits were made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.

In one embodiment, the invention provides a polynucleotide encoding an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein and a serum albumin protein. In a further embodiment, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein and a serum albumin protein. In a preferred embodiment, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein and a serum albumin protein encoded by a polynucleotide described in Table 2. In a further preferred embodiment, the invention provides a polynucleotide encoding an albumin fusion protein whose sequence is shown as SEQ ID NO:Y in Table 2. In other embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a biologically active and/or therapeutically active fragment of a Therapeutic protein and a serum albumin protein. In other embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a biologically active and/or therapeutically active variant of a Therapeutic protein and a serum albumin protein. In preferred embodiments, the serum albumin protein component of the albumin fusion protein is the mature portion of serum albumin. The invention further encompasses polynucleotides encoding these albumin fusion proteins.

In further embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein, and a biologically active and/or therapeutically active fragment of serum albumin. In further embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a Therapeutic protein and a biologically active and/or therapeutically active variant of serum albumin. In preferred embodiments, the Therapeutic protein portion of the albumin fusion protein is the mature portion of the Therapeutic protein. In a further preferred embodiment, the Therapeutic protein portion of the albumin fusion protein is the extracellular soluble domain of the Therapeutic protein. In an alternative embodiment, the Therapeutic protein portion of the albumin fusion protein is the active form of the Therapeutic protein. The invention further encompasses polynucleotides encoding these albumin fusion proteins.

In further embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, a biologically active and/or therapeutically active fragment or variant of a Therapeutic protein and a biologically active and/or therapeutically active fragment or variant of serum albumin. In preferred embodiments, the invention provides an albumin fusion protein comprising, or alternatively consisting of, the mature portion of a Therapeutic protein and the mature portion of serum albumin. The invention further encompasses polynucleotides encoding these albumin fusion proteins.

Therapeutic Proteins

As stated above, a polynucleotide of the invention encodes a protein comprising or alternatively consisting of, at least a fragment or variant of a Therapeutic protein and at least a fragment or variant of human serum albumin, which are associated with one another, preferably by genetic fusion.

An additional embodiment includes a polynucleotide encoding a protein comprising or alternatively consisting of at least a fragment or variant of a Therapeutic protein and at least a fragment or variant of human serum albumin. which are linked with one another by chemical conjugation.

As used herein, “Therapeutic protein” refers to proteins, polypeptides, antibodies, peptides or fragments or variants thereof, having one or more therapeutic and/or biological activities. Therapeutic proteins encompassed by the invention include but are not limited to, proteins, polypeptides, peptides, antibodies, and biologics. (The terms peptides, proteins, and polypeptides are used interchangeably herein.) It is specifically contemplated that the term “Therapeutic protein” encompasses antibodies and fragments and variants thereof. Thus a protein of the invention may contain at least a fragment or variant of a Therapeutic protein, and/or at least a fragment or variant of an antibody. Additionally, the term “Therapeutic protein” may refer to the endogenous or naturally occurring correlate of a Therapeutic protein.

By a polypeptide displaying a “therapeutic activity” or a protein that is “therapeutically active” is meant a polypeptide that possesses one or more known biological and/or therapeutic activities associated with a therapeutic protein such as one or more of the Therapeutic proteins described herein or otherwise known in the art. As a non-limiting example, a “Therapeutic protein” is a protein that is useful to treat, prevent or ameliorate a disease, condition or disorder. As a non-limiting example, a “Therapeutic protein” may be one that binds specifically to a particular cell type (normal (e.g., lymphocytes) or abnormal e.g., (cancer cells)) and therefore may be used to target a compound (drug, or cytotoxic agent) to that cell type specifically.

For example, a non-exhaustive list of “Therapeutic protein” portions which may be comprised by an albumin fusion protein of the invention includes, but is not limited to, erythropoietin (EPO), IL-2, G-CSF, Insulin, Calcitonin, Growth Hormone, IFN-alpha, IFN-beta, PTH, TR6 (International Publication No. WO 98/30694), BLyS, BLyS single chain antibody, Resistin, Growth hormone releasing factor, VEGF-2, KGF-2, D-SLAM, KDI, and TR2, GLP-1, Extendin 4, and GM-CSF.

Interferon hybrids may also be fused to the amino or carboxy terminus of albumin to form an interferon hybrid albumin fusion protein. Interferon hybrid albumin fusion protein may have enhanced, or alternatively, suppressed interferon activity, such as antiviral responses, regulation of cell growth, and modulation of immune response (Lebleu et al., PNAS USA, 73:3107-3111 (1976); Gresser et al., Nature, 251:543-545 (1974); and Johnson, Texas Reports Biol Med, 35:357-369 (1977)). Each interferon hybrid albumin fusion protein can be used to treat, prevent, or ameliorate viral infections (e.g., hepatitis (e.g., HCV); or HIV), multiple sclerosis, or cancer.

In one embodiment, the interferon hybrid portion of the interferon hybrid albumin fusion protein comprises an interferon alpha-interferon alpha hybrid (herein referred to as an alpha-alpha hybrid). For example, the alpha-alpha hybrid portion of the interferon hybrid albumin fusion protein consists, or alternatively comprises, of interferon alpha A fused to interferon alpha D. In a further embodiment, the A/D hybrid is fused at the common BgIII restriction site to interferon alpha D, wherein the N-terminal portion of the A/D hybrid corresponds to amino acids 1-62 of interferon alpha A and the C-terminal portion corresponds to amino acids 64-166 of interferon alpha D. For example, this A/D hybrid would comprise the amino acid sequence: CDLPQTHSLGSRRTLMLLAQMRX1ISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFITKDSSAAWD EDLLDKFCTELYQQLNDLEACVMQEERVGETPLMNX2DSILAVKKYFRRITLYLTEKKYSPCAWEVVRAEIMRSLS LSTNLQERLRRKE (SEQ ID NO:1326), wherein the X1 is R or K and the X2 is A or V (see, for example, Construct ID #2875). In an additional embodiment, the A/D hybrid is fused at the common PvuIII restriction site, wherein the N-terminal portion of the A/D hybrid corresponds to amino acids 1-91 of interferon alpha A and the C-terminal portion corresponds to amino acids 93-166 of interferon alpha D. For example, this A/D hybrid would comprise the amino acid sequence: CDLPQTHSLGSRRTLMLLAQMRX1ISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWD ETLLDKFYTELYQQLNDLEACVMQEERVGETPLMNX2DSILAVKKYFRRITLYLTEKKYSPCAWEVVRAEIMRSLSL STNLQERLRRKE (SEQ ID NO:1311), wherein the X1 is R or K and the second X2 is A or V (see, for example, Construct ID #2872). These hybrids are further described in U.S. Pat. No. 4,414,510, which is hereby incorporated by reference in its entirety.

In an additional embodiment, the alpha-alpha hybrid portion of the interferon hybrid albumin fusion protein consists, or alternatively comprises, of interferon alpha A fused to interferon alpha F. In a further embodiment, the A/F hybrid is fused at the common PvuIII restriction site, wherein the N-terminal portion of the A/F hybrid corresponds to amino acids 1-91 of interferon alpha A and the C-terminal portion corresponds to amino acids 93-166 of interferon alpha F. For example, this A/F hybrid would comprise the amino acid sequence: CDLPQTHSLGSRRTLMLLAQMRXISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWD ETLLDKFYTELYQQLNDMEACVIQEVGVEETPLMNVDSILAVKKYFQRITLYLTEKKYSPCAWEVVRAEIMRSFSL SKIFQERLRRKE (SEQ ID NO:1321), wherein X is either R or K (see, for example, Construct ID #2874). These hybrids are further described in U.S. Pat. No. 4,414,510, which is hereby incorporated by reference in its entirety. In a further embodiment, the alpha-alpha hybrid portion of the interferon hybrid albumin fusion protein consists, or alternatively comprises, of interferon alpha A fused to interferon alpha B. In an additional embodiment, the A/B hybrid is fused at the common PvuIII restriction site, wherein the N-terminal portion of the A/B hybrid corresponds to amino acids 1-91 of interferon alpha A and the C-terminal portion corresponds to amino acids 93-166 of interferon alpha B. For example, this A/B hybrid would comprise an amino acid sequence: CDLPQTHSLGSRRTLMLLAQMRX, ISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWD ETLLDKFYTELYQQLNDLEX2X3X4X5QEVGVIESPLMYEDSILAVRKYFQRITLYLTEKKYSSCAWEVVRAEIMRSFS LSINLQKRLKSKE (SEQ ID NO:1316), wherein the X1 is R or K and X2 through X5 is SCVM or VLCD (see, for example, Construct ID #2873). These hybrids are further described in U.S. Pat. No. 4,414,510, which is hereby incorporated by reference in its entirety.

In another embodiment, the interferon hybrid portion of the interferon hybrid albumin fusion protein comprises an interferon beta-interferon alpha hybrid (herein referred to as a beta-alpha hybrid). For example, the beta-alpha hybrid portion of the interferon hybrid albumin fusion protein consists, or alternatively comprises, of interferon beta-1 fused to interferon alpha D (also referred to as interferon alpha-1). In a further embodiment, the beta-1/alpha D hybrid is fused wherein the N-terminal portion corresponds to amino acids 1-73 of interferon beta-1 and the C-terminal portion corresponds to amino acids 74-167 of interferon alpha D. For example, this beta-1/alpha D hybrid would comprise an amino acid sequence: MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQQFQKEDAALTIYEMLQNIFAIFRQDSSAA WDEDLLDKFCTELYQQLNDLEACVMQEERVGETPLMNXDSILAVKKYFRRITLYLTEKKYSPCAWEVVRAEIMRS LSLSTNLQERLRRKE (SEQ ID NO:2130), wherein X is A or V. These hybrids are further described in U.S. Pat. No. 4,758,428, which is hereby incorporated by reference in its entirety.

In another embodiment, the interferon hybrid portion of the interferon hybrid albumin fusion protein comprises an interferon alpha-interferon beta hybrid (herein referred to as a alpha-beta hybrid). For example, the alpha-beta hybrid portion of the interferon hybrid albumin fusion protein consists, or alternatively comprises, of interferon alpha D (also referred to as interferon alpha-1) fused to interferon beta-1. In a further embodiment, the alpha D/beta-1 hybrid is fused wherein the N-terminal portion corresponds to amino acids 1-73 of interferon alpha D and the C-terminal portion corresponds to amino acids 74-166 of interferon beta-1. For example, this alpha D/beta-1 hybrid would have an amino acid sequence: MCDLPETHSLDNRRTLMLLAQMSRISPSSCLMDRHDFGFPQEEFDGNQFQKAPAISVLHELIQQIFNLFTTKDSSSTG WNETIVENLLANVYHQINHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTIVRVEILRNF YFINRLTGYLRN (SEQ ID NO:2131). These hybrids are further described in U.S. Pat. No. 4,758,428, which is hereby incorporated by reference in its entirety.

In further embodiments, the interferon hybrid portion of the interferon hybrid albumin fusion proteins may comprise additional combinations of alpha-alpha interferon hybrids, alpha-beta interferon hybrids, and beta-alpha interferon hybrids. In additional embodiments, the interferon hybrid portion of the interferon hybrid albumin fusion protein may be modified to include mutations, substitutions, deletions, or additions to the amino acid sequence of the interferon hybrid. Such modifications to the interferon hybrid albumin fusion proteins may be made, for example, to improve levels of production, increase stability, increase or decrease activity, or confer new biological properties.

The above-described interferon hybrid albumin fusion proteins are encompassed by the invention, as are host cells and vectors containing polynucleotides encoding the polypeptides. In one embodiment, a interferon hybrid albumin fusion protein encoded by a polynucleotide as described above has extended shelf life. In an additional embodiment, a interferon hybrid albumin fusion protein encoded by a polynucleotide described above has a longer serum half-life and/or more stabilized activity in solution (or in a pharmaceutical composition) in vitro and/or in vivo than the corresponding unfused interferon hybrid molecule.

In another non-limiting example, a “Therapeutic protein” is a protein that has a biological activity, and in particular, a biological activity that is useful for treating, preventing or ameliorating a disease. A non-inclusive list of biological activities that may be possessed by a Therapeutic protein includes, enhancing the immune response, promoting angiogenesis, inhibiting angiogenesis, regulating endocrine function, regulating hematopoietic functions, stimulating nerve growth, enhancing an immune response, inhibiting an immune response, or any one or more of the biological activities described in the “Biological Activities” section below and/or as disclosed for a given Therapeutic protein in Table 1 (column 2).

As used herein, “therapeutic activity” or “activity” may refer to an activity whose effect is consistent with a desirable therapeutic outcome in humans, or to desired effects in non-human mammals or in other species or organisms. Therapeutic activity may be measured in vivo or in vitro. For example, a desirable effect may be assayed in cell culture. As an example, when EPO is the Therapeutic protein, the effects of EPO on cell proliferation as described in Example 8 may be used as the endpoint for which therapeutic activity is measured. Such in vitro or cell culture assays are commonly available for many Therapeutic proteins as described in the art. Examples of assays include, but are not limited to those described herein in the Examples section or in the “Exemplary Activity Assay” column (column 3) of Table 1.

Therapeutic proteins corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention, such as cell surface and secretory proteins, are often modified by the attachment of one or more oligosaccharide groups. The modification, referred to as glycosylation, can dramatically affect the physical properties of proteins and can be important in protein stability, secretion, and localization. Glycosylation occurs at specific locations along the polypeptide backbone. There are usually two major types of glycosylation: glycosylation characterized by O-linked oligosaccharides, which are attached to serine or threonine residues; and glycosylation characterized by N-linked oligosaccharides, which are attached to asparagine residues in an Asn-X-Ser or Asn-X-Thr sequence, where X can be any amino acid except proline. N-acetylneuramic acid (also known as sialic acid) is usually the terminal residue of both N-linked and 0-linked oligosaccharides. Variables such as protein structure and cell type influence the number and nature of the carbohydrate units within the chains at different glycosylation sites. Glycosylation isomers are also common at the same site within a given cell type.

For example, several types of human interferon are glycosylated. Natural human interferon-α2 is O-glycosylated at threonine 106, and N-glycosylation occurs at asparagine 72 in interferon-α14 (Adolf et al., J. Biochem 276:511 (1991); Nyman T A et al., J. Biochem 329:295 (1998)). The oligosaccharides at asparagine 80 in natural interferon-β1α may play an important factor in the solubility and stability of the protein, but may not be essential for its biological activity. This permits the production of an unglycosylated analog (interferon-β1b) engineered with sequence modifications to enhance stability (Hosoi et al., J. Interferon Res. 8:375 (1988; Karpusas et al., Cell Mol Life Sci 54:1203 (1998); Knight, J. Interferon Res. 2:421 (1982); Runkel et al., Pharm Res 15:641 (1998); Lin, Dev. Biol. Stand. 96:97 (1998)). Interferon-γ contains two N-linked oligosaccharide chains at positions 25 and 97, both important for the efficient formation of the bioactive recombinant protein, and having an influence on the pharmacokinetic properties of the protein (Sareneva et al., Eur. J. Biochem 242:191 (1996); Sareneva et al, Biochem J. 303:831 (1994): Sareneva et al., J. Interferon Res. 13:267 (1993)). Mixed O-linked and N-linked glycosylation also occurs, for example in human erythropoietin, N-linked glycosylation occurs at asparagine residues located at positions 24, 38 and 83 while O-linked glycosylation occurs at a serine residue located at position 126(Lai et al., J. Biol. Chem. 261:3116(1986); Broudy et al., Arch. Biochem. Biophys. 265:329(1988)).

Glycosylation of EPO albumin fusion proteins may influence the activity and/or stability of the EPO albumin fusion proteins. The EPO portion of the albumin fusion protein may contain 3 N-linked sites for glycosylation, each of which can carry one tetra-antennary structure. When the EPO albumin fusion protein is glycosylated, the half-life of the molecule may be increased. In one embodiment, the EPO albumin fusion protein is glycosylated. In another embodiment, the EPO albumin fusion protein is hyperglycosylated.

One type of sugar commonly found in oligosaccharides is sialic acid. Each tetra-antennary structure of the N-linked glycosylation sites of EPO may carry four sialic acid residues. Accordingly, in a preferred embodiment, the EPO albumin fusion protein is glycosylated with a carbohydrate group containing sialic acid. In an additional embodiment, the EPO albumin fusion protein comprises a fully sialylated EPO protein containing four sialic acid residues per tetra-antennerary structure per site with a molar ratio of sialic acid to protein 12:1 or greater. In alternative embodiments, the EPO albumin fusion protein comprises a hypersialylated EPO protein wherein one, two, or three sialic acid residues are attached at each tetra-antennerary structure per site with a molar ratio of sialic acid to protein less than 12:1.

Two types of sialic acid that may be used in the sialylation of the EPO albumin fusion protein are N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc). In a preferred embodiment, hypersialylated EPO albumin fusion proteins contain Neu5Ac. More preferably, the total sialic acid content of hypersialylated EPO albumin fusion proteins is at least 97% Neu5Ac. Most preferred are EPO albumin fusion protein structures with little or no Neu5Gc.

Preferably, the albumin EPO fusion protein has at least 4 moles of sialylation, and more preferably, at least 8-9 moles of sialylation. An additional embodiment comprises an albumin EPO fusion protein with 4 moles of sialylation, 5 moles of sialylation, 6 moles of sialylation, 7 moles of sialylation, 8-9 moles of sialylation, 8 moles of sialylation, 9 moles of sialylation, 10 moles of sialylation, 11 moles of sialylation, or 12 moles of sialylation.

The degree of sialylation of a protein changes the charge of the protein and its retention time on a chromatography column. Therefore, certain chromatography steps used in the purification process may be used to monitor or enrich for hypersialylated EPO albumin fusion proteins. In a preferred embodiment, the amount of sialylation may be monitored by HPLC chromatography. In an additional embodiment, steps in the purification process of EPO albumin fusions may be used to enrich for hypersialylated EPO albumin fusion proteins. In a preferred embodiment the purification steps that may be used to enrich for hypersialylated EPO albumin fusion proteins comprise the butyl-sepharose FF purification step to remove virus particles by high ammonium salt and the hydroxyapatite chromatography at pH 6.8 for the final purification step.

Therapeutic proteins corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention, as well as analogs and variants thereof, may be modified so that glycosylation at one or more sites is altered as a result of manipulation(s) of their nucleic acid sequence, by the host cell in which they are expressed, or due to other conditions of their expression. For example, glycosylation isomers may be produced by abolishing or introducing glycosylation sites, e.g., by substitution or deletion of amino acid residues, such as substitution of glutamine for asparagine, or unglycosylated recombinant proteins may be produced by expressing the proteins in host cells that will not glycosylate them, e.g. in E. coli or glycosylation-deficient yeast. These approaches are described in more detail below and are known in the art.

Therapeutic proteins, particularly those disclosed in Table 1, and their nucleic acid and amino acid sequences are well known in the art and available in public databases such as Chemical Abstracts Services Databases (e.g., the CAS Registry), GenBank, and subscription provided databases such as GenSeq (e.g., Derwent). Exemplary nucleotide sequences of Therapeutic proteins which may be used to derive a polynucleotide of the invention are shown in column 7, “SEQ ID NO:X,” of Table 2. Sequences shown as SEQ ID NO:X may be a wild type polynucleotide sequence encoding a given Therapeutic protein (e.g., either full length or mature), or in some instances the sequence may be a variant of said wild type polynucleotide-sequence (e.g., a polynucleotide which encodes the wild type Therapeutic protein, wherein the DNA sequence of said polynucleotide has been optimized, for example, for expression in a particular species; or a polynucleotide encoding a variant of the wild type Therapeutic protein (i.e., a site directed mutant; an allelic variant)). It is well within the ability of the skilled artisan to use the sequence shown as SEQ ID NO:X to derive the construct described in the same row. For example, if SEQ ID NO:X corresponds to a full length protein, but only a portion of that protein is used to generate the specific CID, it is within the skill of the art to rely on molecular biology techniques, such as PCR, to amplify the specific fragment and clone it into the appropriate vector.

Additional Therapeutic proteins corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention include, but are not limited to, one or more of the Therapeutic proteins or peptides disclosed in the “Therapeutic Protein X” column of Table 1 (column 1), or fragment or variable thereof.

Table 1 provides a non-exhaustive list of Therapeutic proteins that correspond to a Therapeutic protein portion of an albumin fusion protein of the invention, or an albumin fusion protein encoded by a polynucleotide of the invention. The first column, “Therapeutic Protein X,” discloses Therapeutic protein molecules that may be followed by parentheses containing scientific and brand names of proteins that comprise, or alternatively consist of, that Therapeutic protein molecule or a fragment or variant thereof. “Therapeutic protein X” as used herein may refer either to an individual Therapeutic protein molecule, or to the entire group of Therapeutic proteins associated with a given Therapeutic protein molecule disclosed in this column. The “Biological activity” column (column 2) describes Biological activities associated with the Therapeutic protein molecule. Column 3, “Exemplary Activity Assay,” provides references that describe assays which may be used to test the therapeutic and/or biological activity of a Therapeutic protein:X or an albumin fusion protein comprising a Therapeutic protein X (or fragment thereof) portion. Each of the references cited in the “Exemplary Activity Assay” column are herein incorporated by reference in their entireties, particularly with respect to the description of the respective activity assay described in the reference (see Methods section therein, for example) for assaying the corresponding biological activity set forth in the “Biological Activity” column of Table 1. The fourth column, “Preferred Indication: Y,” describes disease, disorders, and/or conditions that may be treated, prevented, diagnosed, and/or ameliorated by Therapeutic protein X or an albumin fusion protein comprising a Therapeutic protein X (or fragment thereof) portion. The “Construct ID” column (column 5) provides a link to an exemplary albumin fusion construct disclosed in Table 2 which encodes an albumin fusion protein comprising, or alternatively consisting of the referenced Therapeutic Protein X (or fragment thereof) portion.

TABLE 1
Therapeutic Therapeutic
Protein: X Biological Activity Exemplary Activity Assay Preferred Indication: Y Construct ID Protein: Z
EPO (Erythropoietin; Stimulates cellular Cell proliferation assay using a Anemia; Anemia in Renal Disease; Anemia in 1772, 1774, 1781, 1783, 1793, See Table 2, SEQ
Epoetin alfa; Epoetin differentiation of bone- erythroleukemic cell line TF-1. Oncology Patients; Bleeding Disorders; Chronic Renal 1794, 1925, 1926, 1966, 1969, ID NO: Z for
beta; Gene-activated marrow stem cells at an (Kitamura et al. 1989 J. Cell. Failure; Chronic Renal Failure in Pre-Dialysis 1980, 1981, 1994, 1995, 1996, particular
erythropoietin; early stage of Physiol. 140: 323) Patients; Renal Disease; End-Stage Renal Disease; 1997, 2047, 2102, 2283, 2284, construct.
Darbepoetin-alpha; erythropoiesis; accelerates End-Stage Renal Disease in Dialysis Patients; 2287, 2289, 2294, 2298, 2310,
NESP; Epogen; the proliferation and Chemotherapy; Chemotherapy in Cancer Patients; 2311, 2325, 2326, 2344, 2363,
Procrit; Eprex; maturation of terminally Anemia in zidovudine-treated HIV patients; Anemia in 2373, 2387, 2414, 2441, 2603,
Erypo; Espo; differentiating cells into zidovudine-treated patients; Anemia in HIV patients; 2604, 2605, 3194, 3195, 3196,
Epoimmun; erythrocytes; and Anemia in premature infants; Surgical patients (pre
EPOGIN; modulates the level of and/or post surgery); Surgical patients (pre and/or post
NEORECORMON; circulating erythrocytes. surgery) who are anemic; Surgical patients (pre and/or
HEMOLINK; post surgery) who are undergoing elective surgery;
Dynepo; ARANESP) Surgical patients (pre and/or post surgery) who are
undergoing elective, non-cardiac surgery; Surgical
patients (pre and/or post surgery) who are undergoing
elective, non-cardiac, non-vascular surgery; Surgical
patients (pre and/or post surgery) who are undergoing
elective, non-vascular surgery; Surgical patients (pre
and/or post surgery) who are undergoing cardiac
and/or vascular surgery; Aplastic anemia; Refractory
anemia; Anemia in Inflammatory Bowel Disease;
Refractory anemia in Inflammatory Bowel Disease;
Transfusion avoidance; Transfusion avoidance for
surgical patients; Transfusion avoidance for elective
surgical patients; Transfusion avoidance for elective
orthopedic surgical patients; Patients who want to
Increase Red Blood Cells.
G-CSF (Granulocyte Stimulates the proliferation Proliferation of murine NFS-60 Chemoprotection; Adjunct to Chemotherapy; 1642, 1643, 2363, 2373, 2387, See Table 2, SEQ
colony-stimulating and differentiation of the cells (Weinstein et al, Proc Natl Inflammatory disorders; Cancer; Leukemia; 2414, 2441, 2702, 2637, 2700, ID NO: Z for
factor; Granulokine; progenitor cells for Acad Sci USA 1986; 83, Myelocytic leukemia; Neutropenia, Primary 2701, 2703, 2886, 2887, 2888, particular
KRN 8601; granulocytes and pp5010-4) neutropenias (e.g.; Kostmann syndrome); Secondary 2889, 2890, construct.
Filgrastim; monocytes-macrophages. neutropenia; Prevention of neutropenia; Prevention
Lenograstim; and treatment of neutropenia in HIV-infected patients;
Meograstim; Prevention and treatment of neutropenia associated
Nartograstim; with chemotherapy; Infections associated with
Neupogen; NOPIA; neutropenias; Myelopysplasia; Autoimmune disorders;
Gran; Psoriasis; Mobilization of hematopoietic progenitor
GRANOCYTE; cells; Wound Healing; Autoimmune Disease;
Granulokine; Transplants; Bone marrow transplants; Acute
Neutrogin; Neu-up; myelogeneous leukemia; Lymphoma, Non-Hodgkin's
Neutromax) lymphoma; Acute lymphoblastic leukemia; Hodgkin's
disease; Accelerated myeloid recovery; Glycogen
storage disease.
GM-CSF Regulates hematopoietic Colony Stimulating Assay: Bone Marrow Disorders; Bone marrow transplant; 1697, 1699, 2066, and 2067. See Table 2, SEQ
(Granulocyte- cell differentiation, gene Testa, N. G., et al., “Assays for Chemoprotection; Hepatitis C; HIV Infections; ID NO: Z for
macrophage colony- expression, growth, and hematopoietic growth factors.” Cancer; Lung Cancer; Melanoma; Malignant particular
stimulating factor; function. Balkwill FR (edt) Cytokines, A melanoma; Mycobacterium avium complex; Mycoses; construct.
rhuGM-CSF; BI practical Approach, pp 229-44; Leukemia; Myeloid Leukemia; Infections; Neonatal
61012; Prokine; IRL Press Oxford 1991. infections; Neutropenia; Mucositis; Oral Mucositis;
Molgramostim; Prostate Cancer; Stem Cell Mobilization; Vaccine
Sargramostim; GM- Adjuvant; Ulcers (such as Diabetic, Venous Stasis, or
CSF/IL 3 fusion; Pressure Ulcers); Prevention of neutropenia; Acute
Milodistim; myelogenous leukemia; Hematopoietic progenitor cell
Leucotropin; mobilization; Lymphoma; Non-Hodgkin's lymphoma;
PROKINE; Acute Lymphoblastic Leukemia; Hodgkin's disease;
LEUKOMAX; Accelerated myeloid recovery; Transplant Rejection;
Interberin; Leukine; Xenotransplant Rejection.
Leukine Liquid;
Pixykine)
Human growth Binds to two GHR Ba/F3-hGHR proliferation Acromegaly; Growth failure; Growth hormone 3163, 2983, See Table 2, SEQ
hormone molecules and Induces assay, a novel specific bioassay replacement; Growth hormone deficiency; Pediatric ID NO: Z for
(Pegvisamont; signal transduction through for serum human growth Growth Hormone Deficiency; Adult Growth Hormone particular
Somatrem; receptor dimerization hormone. J Clin Endocrinol Deficiency; Idiopathic Growth Hormone Deficiency; construct.
Somatropin; Metab 2000 Nov; 85(11): 4274-9 Growth retardation; Prader-Willi Syndrome; Prader-
TROVERT; Plasma growth hormone (GH) Willi Syndrome in children 2 years or older; Growth
PROTROPIN; BIO- immunoassay and tibial deficiencies; Growth failure associated with chronic
TROPIN; bioassay, Appl Physiol 2000 renal insufficiency; Osteoporosis; Postmenopausal
HUMATROPE; Dec; 89(6): 2174-8 osteoporosis; Osteopenia, Osteoclastogenesis; burns;
NUTROPIN; Growth hormone (hGH) Cachexia; Cancer Cachexia; Dwarfism; Metabolic
NUTROPIN AQ; receptor mediated cell mediated Disorders; Obesity; Renal failure; Turner's Syndrome;
NUTROPHIN; proliferation, Growth Horm Fibromyalgia; Fracture treatment; Frailty, AIDS
NORDITROPIN; IGF Res 2000 Oct; 10(5): 248-55 wasting; Muscle Wasting; Short Stature; Diagnostic
GENOTROPIN; International standard for Agents; Female Infertility; lipodystrophy.
SAIZEN; growth hormone, Horm Res
SEROSTIM) 1999; 51 Suppl 1: 7-12
Insulin (Human Stimulates glucose uptake Insulin activity may be assayed Hyperglycemia; Diabetes; Diabetes Insipidus; 2250, 2255, 2276, 2278, 2656, See Table 2, SEQ
insulin; Insulin and promotes glycogenesis in vitro using a [3-H]-glucose Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; 2668, 2669, 2671, 2821, 2822, ID NO: Z for
aspart; Insulin and lipogenesis. uptake assay. (J Biol Chem Insulin resistance; Insulin deficiency; Hyperlipidemia; 2832, 2877, 2878, 2882, 2885, particular
Glargine; Insulin 1999 Oct 22; 274(43): 30864-30873). Hyperketonemia; Non-insulin dependent Diabetes 2891, 2897, 2930, 2931, 2942, construct.
lispro; Lys-B28 Pro- Mellitus (NIDDM); Insulin-dependent Diabetes 2986, 3025, 3133, 3134, 3197,
B29; lyspro; LY Mellitus (IDDM); A Condition Associated With 3198, 2726, 2727, 2784, 2789
275585; Diabetes Including, But Not Limited To Obesity,
diarginylinsulin; Heart Disease, Hyperglycemia, Infections,
Des-B26-B30- Retinopathy, And/Or Ulcers; Metabolic Disorders;
insulin-B25-amide; Immune Disorders; Obesity; Vascular Disorders;
Insulin detemir; Suppression of Body Weight; Suppression of
LABI; NOVOLIN; Appetite; Syndrome X.
NOVORAPID;
HUMULIN;
NOVOMIX 30;
VELOSULIN;
NOVOLOG;
LANTUS; ILETIN;
HUMALOG;
MACRULIN;
EXUBRA;
INSUMAN;
ORALIN;
ORALGEN;
HUMAHALE;
HUMAHALIN)
Interferon alfa Confers a range of cellular Anti-viral assay: Rubinstein S, Viral infections; HIV Infections; Hepatitis; Chronic 2249, 2343, 2366, 2381, 2382, See Table 2, SEQ
(Interferon alfa-2b; responses including Familletti PC, Pestka S. (1981) Hepatitis; Hepatitis B; Chronic Hepatitis B; Hepatitis 2410, and 3165. ID NO: Z for
recombinant; antiviral, antiproliferative, Convenient assay for C; Chronic Hepatitis C; Hepatitis D; Chronic Hepatitis particular
Interferon alfa-n1; antitumor and interferons. J. Virol. 37(2): 755-8; D; Human Papillomavirus; Herpes Simplex Virus construct.
Interferon alfa-n3; immunomodulatory Anti-proliferation assay: Infection; External Condylomata Acuminata; HIV;
Peginterferon alpha- activities; stimulate Gao Y, et al (1999) Sensitivity HIV Infection; Oncology; Cancer; Solid Tumors;
2b; Ribavirin and production of two of an epstein-barr virus- Melanoma; Malignant Melanoma; Renal Cancer (e.g.,
interferon alfa-2b; enzymes: a protein kinase positive tumor line, Daudi, to Renal Cell Carcinoma); Lung Cancer (e.g,. Non-Small
Interferon alfacon-1; and an oligoadenylate alpha interferon correlates with Cell Lung Cancer or Small Cell Lung Cancer) Colon
interferon consensus; synthetase. expression of a GC-rich viral Cancer; Breast Cancer; Liver Cancer; Prostate Cancer;
YM 643; CIFN; transcript. Mol Cell Biol. Bladder Cancer; Gastric Cancer; Sarcoma; AIDS-
interferon-alpha 19(11): 7305-13. Related Kaposi's Sarcoma; Lymphoma; T Cell
consensus; Lymphoma; Cutaneous T-Cell Lymphoma; Non-
recombinant Hodgkin's Lymphoma; Brain Cancer; Glioma;
methionyl consensus Glioblastoma Multiforme; Cervical Dysplasia;
interferon; Leukemia; Preleukemia; Bone Marrow Disorders;
recombinant Bone Disorders; Hairy Cell Leukemia; Chronic
consensus interferon; Myelogeonus Leukemia; Hematological Malignancies;
CGP 35269; RO Hematological Disorders; Multiple Myeloma;
253036; RO 258310; Bacterial Infections; Chemoprotection;
INTRON A; PEG- Thrombocytopenia; Multiple Sclerosis; Pulmonary
INTRON; OIF; Fibrosis; Age-Related Macular Degeneration; Macular
OMNIFERON; Degeneration; Crohn's Disease; Neurological
PEG-OMNIFERON; Disorders; Arthritis; Rheumatoid Arthritis; Ulcerative
VELDONA; PEG- Colitis; Osteoporosis, Osteopenia, Osteoclastogenesis;
REBETRON; Fibromyalgia; Sjogren's Syndrome; Chronic Fatigue
ROFERON A; Syndrome; Fever; Hemmorhagic Fever; Viral
WELLFERON; Hemmorhagic Fevers; Hyperglycemia; Diabetes;
ALFERON N/LDO; Diabetes Insipidus; Diabetes mellitus; Type 1 diabetes;
REBETRON; Type 2 diabetes; Insulin resistance; Insulin deficiency;
ALTEMOL; Hyperlipidemia; Hyperketonemia; Non-insulin
VIRAFERONPEG; dependent Diabetes Mellitus (NIDDM); Insulin-
PEGASYS; dependent Diabetes Mellitus (IDDM); A Condition
VIRAFERON; Associated With Diabetes Including, But Not Limited
VIRAFON; To Obesity, Heart Disease, Hyperglycemia, Infections,
AMPLIGEN; Retinopathy, And/Or Ulcers; Metabolic Disorders;
INFERGEN; Immune Disorders; Obesity; Vascular Disorders;
INFAREX; Suppression of Body Weight; Suppression of
ORAGEN) Appetite; Syndrome X.
Calcitonin (Salmon Regulates levels of calcium Hypocalcemic Rat Bioassay, Bone Disorders; Fracture prevention; Hypercalcemia; 1833, 1834, 1835, 1836, 2447, See Table 2, SEQ
Calcitonin and phosphate in serum; bone resorbing assay and the Malignant hypercalcemia; Osteoporosis; Paget's 2513, 2806, 2915 ID NO: Z for
(Salcatonin); causes a reduction in serum pit assay, CT receptor binding disease; Osteopenia, Osteoclastogenesis; osteolysis; particular
Calcitonin human- calcium - an effect opposite assay, CAMP stimulation osteomyelitis; osteonecrosis; periodontal bone loss; construct.
salmon hybrid; to that of human assay: J Bone Miner Res 1999 osteoarthritis; rheumatoid arthritis; osteopetrosis;
Forcaltonin; Fortical; parathyroid hormone. Aug; 14(8): 1425-31 periodontal, lytic, or metastatic bone disease;
Calcitonin; Calcitonin osteoclast differentiation inhibition; bone disorders;
a Almirall; bone healing and regeneration.
Calcitonina Hubber;
Calcimar; Calsynar;
Calogen; Miacalcic;
Miacalcin;
SB205614;
Macritonin;
Cibacalcin;
Cibacalcina;
Cibacalcine;
Salmocalcin;
PowderJect
Calcitonin)
(CAS-21215-62-3)
Interferon beta Modulates MHC antigen Anti-viral assay: Rubinstein S, Multiple Sclerosis; Oncology; Cancer; Solid Tumors; 1778, 1779, 2011, 2013, 2053, See Table 2, SEQ
(Interferon beta-1a; expression, NK cell activity Familletti PC, Pestka S. (1981) Melanoma; Malignant Melanoma; Renal Cancer (e.g., 2054, 2492, 2580, 2795, 2796, ID NO: Z for
Interferon beta 1b; and IFNg production and Convenient assay for Renal Cell Carcinoma); Lung Cancer (e.g,. Non-Small 2797. particular
Interferon-beta- IL12 production in interferons. J. Virol. 37(2): 755-8; Cell Lung Cancer or Small Cell Lung Cancer) Colon construct.
serine; SH 579; ZK monocytes. Anti-proliferation assay: Cancer; Breast Cancer; Liver Cancer; Prostate Cancer;
157046; BCDF; Gao Y, et al (1999) Sensitivity Bladder Cancer; Gastric Cancer; Sarcoma; AIDS-
beta-2 IF; Interferon- of an epstein-barr virus- Related Kaposi's Sarcoma; Lymphoma; T Cell
beta-2; rhIL-6; positive tumor line, Daudi, to Lymphoma; Cutaneous T-Cell Lymphoma; Non-
SJ0031; DL 8234; alpha interferon correlates with Hodgkin's Lymphoma; Brain Cancer; Glioma;
FERON; IFNbeta; expression of a GC-rich viral Glioblastoma Multiforme; Cervical Dysplasia;
BETASERON; transcript. Mol Cell Biol. Leukemia; Preleukemia; Bone Marrow Disorders;
AVONEX; REBIF; 19(11): 7305-13. Bone Disorders; Hairy Cell Leukemia; Chronic
BETAFERON; Myelogeonus Leukemia; Hematological Malignancies;
SIGOSIX) Hematological Disorders; Multiple Myeloma;
Bacterial Infections; Chemoprotection;
Thrombocytopenia; Viral infections; HIV Infections;
Hepatitis; Chronic Hepatitis; Hepatitis B; Chronic
Hepatitis B; Hepatitis C; Chronic Hepatitis C;
Hepatitis D; Chronic Hepatitis D; Human
Papillomavirus; Herpes Simplex Virus Infection;
External Condylomata Acuminata; HIV; HIV
Infection; Pulmonary Fibrosis; Age-Related Macular
Degeneration; Macular Degeneration; Crohn's
Disease; Neurological Disorders; Arthritis;
Rheumatoid Arthritis; Ulcerative Colitis;
Osteoporosis, Osteopenia, Osteoclastogenesis;
Fibromyalgia; Sjogren's Syndrome; Chronic Fatigue
Syndrome; Fever; Hemmorhagic Fever; Viral
Hemmorhagic Fevers; Hyperglycemia; Diabetes;
Diabetes Insipidus; Diabetes mellitus; Type 1 diabetes;
Type 2 diabetes; Insulin resistance; Insulin deficiency;
Hyperlipidemia; Hyperketonemia; Non-insulin
dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM); A Condition
Associated With Diabetes Including, But Not Limited
To Obesity, Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Growth hormone Acts on the anterior Growth hormone-releasing Acromegaly; Growth failure; Growth hormone 1747 and 1748. See Table 2, SEQ
releasing factor; pituitary to stimulate the peptides (GHRPs) are known to replacement; Growth hormone deficiency; Pediatric ID NO: Z for
Growth hormone production and secretion of release growth hormone (GH) Growth Hormone Deficiency; Adult Growth Hormone particular
releasing hormone growth hormone and exert in vivo and in vitro by a direct Deficiency; Idiopathic Growth Hormone Deficiency; construct.
(Sermorelin acetate; a trophic effect on the action on receptors in anterior Growth retardation; Prader-Willi Syndrome; Prader-
Pralmorelin; gland. pituitary cells. Biological Willi Syndrome in children 2 years or older; Growth
Somatorelin; activity can be measured in cell deficiencies; Growth failure associated with chronic
Somatoliberin; lines expressing growth renal insufficiency; Osteoporosis; Osteopenia,
Geref; Gerel; hormone releasing factor Osteoclastogenesis; Postmenopausal osteoporosis;
Groliberin) receptor (Mol Endocrinol 1992 burns; Cachexia; Cancer Cachexia; Dwarfism;
Oct; 6(10): 1734-44, Molecular Metabolic Disorders; Obesity; Renal failure; Turner's
Endocrinology, Vol 7, 77-84). Syndrome; Fibromyalgia; Fracture treatment; Frailty,
AIDS wasting; Muscle Wasting; Short Stature;
Diagnostic Agents; Female Infertility; lipodystrophy.
IL-2 (Aldesleukin; Promotes the growth of B T cell proliferation assay Cancer; Solid Tumors; Metastatic Renal Cell 1757, 1758, 1812, 1813, 1952, See Table 2, SEQ
interleukin-2 fusion and T cells and augments “Biological activity of Carcinoma; Metastatic Melanoma; Malignant 1954, 2030, and 2031. ID NO: Z for
toxin; T cell growth NK cell and CTL cell recombinant human Melanoma; Melanoma; Renal Cell Carcinoma; Renal particular
factor; killing activity. interleukin-2 produced in Cancer; Lung Cancer (e.g,. Non-Small Cell Lung construct.
PROLEUKIN; Escherichia coli.” Science 223: Cancer or Small Cell Lung Cancer); Colon Cancer;
IMMUNACE; 1412-1415, 1984. natural Breast Cancer; Liver Cancer; Leukemia; Preleukemia;
CELEUK; killer (NK) cell and CTL Hematological Malignancies; Hematological
ONCOLIPIN 2; cytotoxicity assay “Control of Disorders; Acute Myeloid Leukemia; Melanoma;
MACROLIN) homeostasis of CD8+ memory Malignant Melanoma; Non-Hodgkin's Lymphoma;
T cells by opposing cytokines. Ovarian Cancer; Prostate Cancer; Brain Cancer;
Science 288: 675-678, 2000; Glioma; Glioblastoma Multiforme; Hepatitis; Hepatitis
CTLL-2 Proliferation: Gillis et C; Lymphoma; HIV Infection (AIDS); Inflammatory
al (1978) J. Immunol. 120, Bowel Disorders; Kaposi's Sarcoma; Multiple
2027 Sclerosis; Arthritis; Rheumatoid Arthritis; Transplant
Rejection; Diabetes; Type 1 Diabetes Mellitus; Type 2
Diabetes.
Parathyroid Acts in conjuction with Adenylyl cyclase stimulation in Bone Disorders; Fracture prevention; Hypercalcemia; 1749, 1750, 1853, 1854, 1889, See Table 2, SEQ
hormone; parathyrin calcitonin to control rat osteosarcoma cells, Malignant hypercalcemia; Osteoporosis; Paget's 1906, 1907, 1914, 1932, 1938, ID NO: Z for
(PTH; Ostabolin; calcium and phosphate ovariectomized rat model of disease; Osteopenia, Osteoclastogenesis; osteolysis; 1941, 1949, 2021, 2022, 2023, particular
ALX1-11; hPTH 1-34; metabolism; elevates blood osteoporosis: IUBMB Life osteomyelitis; osteonecrosis; periodontal bone loss; 2428, 2714, 2791, 2965, 2966. construct.
LY 333334; MN calcium level; stimulates 2000 Feb; 49(2): 131-5 osteoarthritis; rheumatoid arthritis; osteopetrosis;
10T; parathyroid the activity of osteocytes; periodontal, lytic, or metastatic bone disease;
hormone (1-31); enhances absorption of osteoclast differentiation inhibition; bone disorders;
FORTEO; Ca+/Pi from small intestine bone healing and regeneration.
PARATHAR) into blood; promotes
reabsorption of Ca+ and
inhibits Pi by kidney
tubules.
Resistin Mediates insulin resistance Ability of resistin to influence Hyperglycemia; Diabetes; Diabetes Insipidus; 2295, 2296, 2297, 2300, and See Table 2, SEQ
in Type II diabetes; type II diabetes can be Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; 2309. ID NO: Z for
inhibits insulin-stimulated determined using assays Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
glucose uptake known in the art: Pontoglio et Hyperketonemia; Non-insulin dependent Diabetes construct.
al., J Clin Invest 1998 May Mellitus (NIDDM); Insulin-dependent Diabetes
15; 101(10): 2215-22. Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
TR6 (DcR3; Inhibits Fas Ligand and Cellular apoptosis can be Fas Ligand or LIGHT induced apoptotic disorders: 1520, 1537, 1545, 1546, See Table 2,
Decoy Receptor 3; AIM-2 (TL5, LIGHT) measured by annexin hepatitis; liver failure (including fulminant liver 1568, 1570, 1622, 1623, SEQ ID NO: Z
FASTR) mediated apoptosis. staining, TUNEL staining, failure); graft versus host disease; graft rejection; 1645, 1700, 1702, 1703, for particular
measurement of caspase myelodysplastic syndrome; renal failure; insulin 1704, 1891, 1892, 1912, and construct.
levels. Inhibition of cell dependent diabetes mellitus; rheumatoid arthritis; 1913.
growth can also be directly inflammatory bowel disease; autoimmune disease;
measured, for example by toxic epidermal necrolysis; multiple sclerosis.
ALOMAR Blue staining.
Assay refs: cytotoxicity assay
on human fibrosarcoma
(Epsevik and Nissen-Meyer,
1986, J. Immunol. methods).
DeCAF (D-SLAM; Inhibits proliferation and DeCAF activity can be B cell and/or T cell mediated immune disorders; 1657. See Table 2,
BCM-like differentiation of B cells; determined using assays Immunodeficiency (e.g., Common Variable SEQ ID NO: Z
membrane protein; Antagonize BLyS known in the art, such as for Immunodeficiency, Selective IgA Deficiency) for particular
BLAME (B activity example, those described in construct.
lymphocyte Examples 32-33 of
activator International Publication No.
macrophage WO0111046.
expressed))
BLyS (B Promotes proliferation, BLyS activity can be B cell and/or T cell mediated immune disorders, 1680, 2095, and 2096. See Table 2,
Lymphocyte differentiation and determined using assays particularly immune system disorders associated SEQ ID NO: Z
Stimulator; survival of B cells; known in the art, such as, for with low B cell numbers or low serum for particular
Neutrokine alpha; Promotes example, the costimulatory immunoglobulin; Immunodeficiency (e.g., Common construct.
TL7; BAFF; immunoglobulin proliferation assay and other Variable Immunodeficiency, Selective IgA
TALL-1; THANK; production by B cells. assays disclosed by Moore et Deficiency). Radiolabeled forms: lymphoma, non-
radiolabeled al., 1999, Science, Hodgkins lymphoma, chronic lymphocytic
BLyS) 285(5425): 260-3. leukemia, multiple myeloma.
Anti-BLyS single Agonize or antagonize BLyS agonist or antagonist B cell and/or T cell mediated immune disorders; 1821, 1956, 2501, 2502, See Table 2,
chain antibody (scFvI116A01, BlyS activity. activity can be determined Autoimmune disorders, particularly autoimmune 2638. SEQ ID NO: Z.
scFvI050B11, using assays known in the diseases associated with the production of for particular
scFvI006D08) and art, such as, for example, a autoantibodies; Rheumatoid Arthritis, Systemic construct.
others. modified version the Lupus Erythmatosus; Sjogren's Syndrome, cancers
costimulatory proliferation expressing Blys as an autocrine growth factor, e.g.
assay disclosed by Moore et certain chronic lymphocytic leukemias.
al., 1999, Science,
285(5425): 260-3, in which
BlyS is mixed or
preincubated with the anti-
BlyS antibody prior to being
applied to the responder B
lymphocytes.
MPIF-1 (Myeloid Inhibits myeloid MPIF-1 activity can be Chemoprotection; Adjunct to Chemotherapy; 1681, 3166, 3167, 3168, See Table 2,
Progenitor progenitor cells; and measured using the Inflammatory disorders; Cancer; Leukemia; SEQ ID NO: Z
Inhibitory Factor; activates monocytes myeloprotection assay and Myelocytic leukemia; Neutropenia, Primary for particular
CK beta-8; chemotaxis assay described neutropenias (e.g.; Kostmann syndrome); Secondary construct.
Mirostipen) in US patent 6,001,606. neutropenia; Prevention of neutropenia; Prevention
and treatment of neutropenia in HIV-infected
patients; Prevention and treatment of neutropenia
associated with chemotherapy; Infections associated
with neutropenias; Myelopysplasia; Autoimmune
disorders; Psoriasis; Mobilization of hematopoietic
progenitor cells; Wound Healing; Autoimmune
Disease; Transplants; Bone marrow transplants;
Acute myelogeneous leukemia; Lymphoma, Non-
Hodgkin's lymphoma; Acute lymphoblastic
leukemia; Hodgkin's disease; Accelerated myeloid
recovery; Glycogen storage disease.
KDI (Keratinocyte Inhibits bone marrow KDI activity can be measured Multiple sclerosis; Hepatitis; Cancer; Viral 1746. See Table 2,
Derived Interferon; proliferation; and shows using the antiviral and cell infections, HIV infections, Leukemia. SEQ ID NO: Z
Interferon Kappa antiviral activity. proliferation assays described for particular
Precursor) in Examples 57-63 of construct.
International Publication No.
WO0107608.
TNFR2 (p75) Binds both TNFa and T-cell proliferation can be Autoimmune disease; Rheumatoid Arthritis; 1777 and 1784. See Table 2,
(ENBREL) TNFb; mediates T-cell measured using assays Psoriatic arthritis; Still's Disease; Ankylosing SEQ ID NO: Z
proliferation by TNF; known in the art. For Spondylitis; Cardiovascular Diseases; Vasulitis; for particular
reduces signs and example, “Lymphocytes: a Wegener's granulomatosis; Amyloidosis; Systemic construct.
structural damage in practical approach” edited Lupus Erythematosus, Insulin-Dependent Diabetes
patients with moderately by: SL Rowland, AJ McMichael - Mellitus; Immunodeficiency Disorders; Infection;
to severly active chapter 6, Inflammation; Inflammatory Bowel Disease;
rheumatoid arthritis pages 138-160 Oxford Chrohn's Disease; Psoriasis; AIDS; Graft Rejection;
(RA). University Press (2000); and Graft Versus Host Disease.
“Current Protocols on CD-
ROM” section 3.12
Proliferation Assays for T-
cell Function John Wiley &
Soncs, Inc. (1999).
Keratinocyte Stimulates epithelial cell KGF-2 activity can be Stimulate Epithelial Cell Proliferation; Stimulate 1785, 1786, 1916, 1917, See Table 2,
growth factor 2 growth. measured using the wound Basal Keratinocytes; Wound Healing; Stimulate 2498, 2499, 2552, 2553, SEQ ID NO: Z
(Repifermin; KGF- healing assays and epithelial Hair Follicle Production; Healing Of Dermal 2584, 2607, 2608, 2606, for particular
2; Fibroblast cell proliferation assays Wounds. Wound Healing; Eye Tissue Wounds, 2630 construct.
Growth Factor-10; described in US patent Dental Tissue Wounds, Oral Cavity Wounds,
FGF-10) 6,077,692. Diabetic Ulcers, Dermal Ulcers, Cubitus Ulcers,
Arterial Ulcers, Venous Stasis Ulcers, Burns
Resulting From Heat Exposure Or Chemicals, or
Other Abnormal Wound Healing Conditions such as
Uremia, Malnutrition, Vitamin Deficiencies or
Complications Associated With Systemic Treatment
With Steroids, Radiation Therapy or Antineoplastic
Drugs or Antimetabolites; Promote Dermal
Reestablishment Subsequent To Dermal Loss;
Increase the Adherence Of Skin Grafts To A Wound
Bed; Stimulate Re-Epithelialization from The
Wound Bed; To Promote Skin Strength; Improve
The Appearance Of Aged Skin; Proliferate
Hepatocytes, Lung, Breast, Pancreas, Stomach,
Bladder, Small Intestine, Large Intestine; Sebocytes,
Hair Follicles, Type II Pneumocytes, Mucin-
Producing Goblet Cells, or Other Epithelial Cells,
Endothelial Cells, Keratinocytes, or Basal
Keratinocytes (and Their Progenitors) Contained
Within The Skin, Lung, Liver, Bladder, Eye,
Salivary Glands, or Gastrointestinal Tract; Reduce
The Side Effects Of Gut Toxicity That Result From
Radiation, Chemotherapy Treatments Or Viral
Infections; Cytoprotector, especially of the Small
Intestine Mucosa or Bladder; Mucositis (Mouth
Ulcers); Regeneration Of Skin; Full and/or Partial
Thickness Skin Defects, including Burns, (e.g.,
Repopulation Of Hair Follicles, Sweat Glands, And
Sebaceous Glands); Psoriasis; Epidermolysis
Bullosa; Blisters; Gastric and/or Doudenal Ulcers;
Reduce Scarring; Inflamamatory Bowel Diseases;
Crohn's Disease; Ulcerative Colitis; Gut Toxicity;
Lung Damage; Repair Of Alveoli And/or Brochiolar
Epithelium; Acute Or Chronic Lung Damage;
Emphysema, ARDS; Inhalation Injuries; Hyaline
Membrane Diseases; Infant Respiratory Distress
Syndrome; Bronchopulmonary Displasia In
Premature Infants; Fulminant Liver Failure;
Cirrhosis, Liver Damage caused by Viral Hepatitis
and/or Toxic Substances; Diabetes Mellitus;
Inflammation.
TR2 (and TR2sv1, Inhibits B cell Co-stimulation B-cell Herpes; immune disorders; autoimmune disease; 1788 and 2129. See Table 2,
TR2SV2; proliferation, and proliferation assay and Ig graft versus host disease; graft rejection; variable SEQ ID NO: Z
TNFRSF14; mediates and inhibits production assay (Moore et immunodeficiency; immunodeficiency syndromes; for particular
HVEM; Herpes Herpes Simplex Virus al., 1999, Science, cancer. construct.
Virus Entry (HSV) infection. 285(5425): 260-3.). HSV-1
Mediator; ATAR) and HSV-2 Infectivity Assay:
International Publication
No. WO 97/04658
Macrophage Chemotactic for Chemokine activities can be Inflammatory diseases; wound healing; 1809, 2137, 2474, 2475, See Table 2,
derived monocyte-derived determined using assays angiogenesis; AIDS infection. 2476, and 2477. SEQ ID NO: Z
chemokine, MDC dendritic cells and IL-2- known in the art: Methods in for particular
(Ckbeta-13) activated natural killer Molecular Biology, 2000, construct.
cells. vol. 138: Chemokine
Protocols. Edited by: A. E. I. Proudfoot,
T. N. C. Wells, and
C. A. Power. © Humana
Press Inc., Totowa, NJ
HAGDG59 Activates MIP1 a release Dendritic cell assays are well Immune disorders; cancer; viral infection; 1830 and 1831. See Table 2,
(Retinal short- in Dendritic Cells. known in the art. For inflammation; sepsis; arthritis; asthma. SEQ ID NO: Z
chain example, J. Immunol. for particular
dehydrogenase) 158: 2919-2925 (1997); J. construct.
Leukoc Biol. 65: 822-828
(1999).
GnRH Promotes release of GnRH is known to cause the Infertility; Kallmann's syndrome or other forms of 1862 and 1863. See Table 2,
(Gonadotropin follicle-stimulating release of follicle stimulating hypergonadotropic hypergonadism (failure to go SEQ ID NO: Z
Releasing hormone and luteinizing hormone (FSH) and/or through puberty naturally). for particular
Hormone) hormone from anterior luteinizing hormone (LH) in construct.
pituitary. vivo by a direct action on
receptors in anterior pituitary
gonadotropes. GnRH activity
can be determined by
measuring FSH levels in the
medium of cultured
gonadotropes before and
after GnRH supplementation.
For example, Baker et al.
Biol Reprod 2000
Sep; 63(3): 865-71.
Teprotide Inhibits angiotensin Inhibition of ACE can be Hypertension; congestive heart failure. 1866, 1867, 2025, and 2026. See Table 2,
converting enzyme determined using assays SEQ ID NO: Z
(ACE). known in the art. For for particular
example, Anzenbacherova et construct.
al., J. Pharma Biomed Anal
2001 Mar; 24(5-6): 1151-6.
Human chemokine Involved in Chemokine activities can be Autoimmune disorders; Immunity; Vascular and 1933, 1934, 1947, 1948, See Table 2,
HCC-1 (ckBeta-1; inflammation, allergy, determined using assays Inflammatory disorders; HIV; AIDS; infectious 1955, 1998, 2355, 2412, SEQ ID NO: Z
HWFBD) tissue rejection, viral known in the art: Methods in diseases. 2449, 2837, 2838, 2839, for particular
infection, and tumor Molecular Biology, 2000, 2840, 2841, 2842, 2843, construct.
biology; enhances vol. 138: Chemokine 2844, 2845, 2849, 2947,
proliferation of CD34+ Protocols. Edited by: A. E. I. Proudfoot 3066, 3105, 3124, 3125,
myeloid progenitor cells. Proudfoot, T. N. C. Wells, and 3139, 3152, 3153, 3154,
C. A. Power. © Humana 3155, 3156, 3169, 3170,
Press Inc., Totowa, NJ 3202, 3203, 3204, 3205,
3206, 3207, 3272
ACE2 inhibitor Inhibits production of Inhibition of angiotensin can Treatment for elevated angiotensin II and/or 1989, 2000, 2001, and 2002. See Table 2,
(DX512) angiotensin II which be determined using assays aldosterone levels, which can lead to SEQ ID NO: Z
induces aldosterone known in the art. For vasoconstriction, impaired cardiac output and/or for particular
production, arteriolar example, in vitro using a hypertension; Cardiovascular Disease; Cardiac construct.
smooth muscle proliferation assay with rat Failure; Diabetes; Type II Diabetes; Proteinuria;
vasoconstriction, and cardiac fibroblasts as Renal disorders, congestive heart failure.
proliferation of cardiac described in Naunyn
fibroblasts, Induces Schmiedebergs Arch
angiogenesis; an enzyme Pharmacol 1999
that converts angiotensin May; 359(5): 394-9.
I to angiotensin 1-9; also
cleaves des-Arg,
bradykinin and
neurotensin.
TR1 (OCIF; Inhibits Coculture Assay for Osteoporosis; Paget's disease; osteopenia; 2016, 2017, 2085, 2086, See Table 2,
Osteoclastogenesis osteoclastogenesis and Osteoclastogenesis, Bone osteolysis; osteomyelitis; osteonecrosis; periodontal 2529, 2530, 2531, 2532, SEQ ID NO: Z
inhibitory factor; bone resorption, and resorption assay using fetal bone loss; osteoarthritis; rheumatoid arthritis; 2555, 2556, 2557, and 2558. for particular
osteoprotegerin, induces fibroblast long-bone organ culture osteopetrosis; periodontal, lytic, or metastatic bone construct.
OPG; tumor proliferation. system, dentine resorption disease; osteoclast differentiation inhibition; bone
necrosis factor assay, and fibroblast disorders; bone healing and regeneration; organ
receptor proliferation assays are each calcification; vascular calcification.
superfamily described in Kwon et al.,
member 11B FASEB J. 12: 845-854
precursor;) (1998).
Human chemokine Chemotactic for both Chemokine activities can be Cancer; Wound healing; Inflammatory disorders; 2101, 2240, 2241, 2245, See Table 2,
Ckbeta-7 activated (CD3+) T cells determined using assays Immmunoregulatory disorders; Atherosclerosis; 2246, 2247, and 2248. SEQ ID NO: Z
and nonactivated (CD14−) known in the art: Methods in Parasitic Infection; Rheumatoid Arthritis; Asthma; for particular
lymphocytes and Molecular Biology, 2000, Autoimmune disorders. construct.
(CD4+) and (CD8+) T vol. 138: Chemokine
lymphocytes and Protocols. Edited by: A. E. I. Proudfoot,
(CD45RA+) T cells T. N. C. Wells, and
C. A. Power. © Humana
Press Inc., Totowa, NJ
CKbeta4 Attracts and activates Chemokine activities can be Cancer; Solid Tumors; Chronic Infection; 2141, 2330, 2335, 2336, See Table 2,
(HGBAN46; microbicidal leukocytes; determined using assays Autoimmune Disorders; Psoriasis; Asthma; Allergy; 2337, 2338, and 2348. SEQ ID NO: Z
HE9DR66) Attracts CCR6- known in the art: Methods in Hematopoiesis; Wound Healing; Bone Marrow for particular
expressing immature Molecular Biology, 2000, Failure; Silicosis; Sarcoidosis; Hyper-Eosinophilic construct.
dendritic cells and vol. 138: Chemokine Syndrome; Lung Inflammation; Fibrotic Disorders;
memory/effector T cells; Protocols. Edited by: A. E. I. Proudfoot, Atherosclerosis; Periodontal diseases; Viral diseases;
B-cell chemotaxis; T. N. C. Wells, and Hepatitis.
inhibits proliferation of C. A. Power. © Humana
myeloid progenitors; Press Inc., Totowa, NJ
chemotaxis of PBMC's.
Leptin Controls obesity through in vivo modulation of food Hyperglycemia; Diabetes; Diabetes Insipidus; 2146, 2184, 2186, and 2187. See Table 2,
regulation of appetite, intake, reduction in body Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; SEQ ID NO: Z
reduction of body weight, and lowering of Insulin resistance; Insulin deficiency; for particular
weight, and lowering of insulin and glucose levels in Hyperlipidemia; Hyperketonemia; Non-insulin construct.
insulin and glucose level. ob/ob mice, dependent Diabetes Mellitus (NIDDM); Insulin-
radioimmunoassay (RIA) and dependent Diabetes Mellitus (IDDM); a Condition
activation of the leptin Associated With Diabetes Including, But Not
receptor in a cell-based Limited To Obesity, Heart Disease, Hyperglycemia,
assay. Protein Expr Purif Infections, Retinopathy, And/Or Ulcers; Metabolic
1998 Dec; 14(3): 335-42 Disorders; Immune Disorders; Obesity; Vascular
Disorders; Suppression of Body Weight;
Suppression of Appetite; Syndrome X;
Immunological Disorders; Immunosuppression.
IL-1 receptor Binds IL1 receptor 1) Competition for IL-1 Autoimmune Disease; Arthritis; Rheumatoid 2181, 2182, 2183, and 2185. See Table 2,
antagonist without activating the binding to IL-1 receptors in Arthritis; Asthma; Diabetes; Diabetes Mellitus; SEQ ID NO: Z
(Anakinra; soluble target cells; inhibits the YT-NCI or C3H/HeJ cells GVHD; Inflammatory Bowel Disorders; Chron's for particular
interleukin-1 binding of IL1-alpha and (Carter et al., Nature 344: Disease; Ocular Inflammation; Psoriasis; Septic construct.
receptor; IRAP; IL1-beta; and neutralizes 633-638, 1990); Shock; Transplant Rejection; Inflammatory
KINERET; the biologic activity of 2) Inhibition of IL-1- Disorders; Rheumatic Disorders; Osteoporosis;
ANTRIL) IL1-alpha and IL1-beta. induced endothelial cell- Postmenopausal Osteoporosis; Stroke.
leukocyte adhesion (Carter et
al., Nature 344: 633-638,
1990);
3) Proliferation assays on
A375-C6 cells, a human
melanoma cell line highly
susceptible to the
antiproliferative action of IL-
1 (Murai T et al., J. Biol.
Chem. 276: 6797-6806,
2001).
TREM-1 Mediates activation of Secretion of cytokines, Inflammation; Sepsis; bacterial infection; 2226 and 2230. See Table 2,
(Triggering neutrophil and chemokines, degranulation, autoimmune diseases; GVHD. SEQ ID NO: Z
Receptor monocytes; Stimulates and cell surface activation for particular
Expressed on neutrophil and markers can be determined construct.
Monocytes 1) monocyte-mediated using assays described in
inflammatory response; Bouchon et al, J Immunol
Promotes secretion of 2000 May 15; 164(10): 4991-5.
TNF, IL-8, and MCP-1;
Induces neutrophil
degranulation, Ca2+
mobilization and tyrosine
phosphorylation of
extracellular signal-
related kinase 1 (ERK1),
ERK2 and phospholipase
C-gamma.
HCNCA73 Induces T-cell activation- FMAT can be used to Autoimmune disorders; Inflammation of the 2244 and 2365. See Table 2,
expression of CD152 measure T-cell surface gastrointestinal tract; Cancer; Colon Cancer; SEQ ID NO: Z
marker; Stimulates markers (CD69, CD152, Allergy; Crohn's disease. for particular
release of TNF-a and CD71, HLA-DR) and T-cell construct.
MIP-1a from immature, cytokine production (e.g.,
monocyte-derived IFNg production). J. of
dendritic cells; Promotes Biomol. Screen. 4: 193-204
maturation of dendritic (1999). Other T-cell
cells. proliferation assays:
“Lymphocytes: a practical
approach” edited by: SL Rowland,
AJ McMichael -
Chapter 6, pages 138-160
Oxford University Press
(2000); WO 01/21658
Examples 11-14, 16-17 and
33.
VEGF-2 (Vascular Promotes endothelial cell VEGF activity can be Coronary artery disease; Critical limb ischemia; 2251, 2252, 2256, and 2257. See Table 2,
Endothelial proliferation. determined using assays Vascular disease; proliferation of endothelial cells, SEQ ID NO: Z
Growth Factor-2; known in the art, such as both vascular and lymphatic. Antagonists may be for particular
VEGF-C) those disclosed in useful as anti-angiogenic agents; Cancer. construct.
International Publication No.
WO0045835, for example.
HCHNF25 Activates MIP1a Release Dendritic cell assays are well Immune disorders; cancer. 2271, 2280, and 2320. See Table 2,
(jumping in Dendritic Cells. known in the art. For SEQ ID NO: Z
translocation example, J. Immunol. for particular
breakpoint) 158: 2919-2925 (1997); J. construct.
Leukoc. Biol. 65: 822-828
(1999).
HLDOU18 (Bone Activates L6/GSK3 Assays for activation of Hyperglycemia; Diabetes; Diabetes Insipidus; 22328, 2340, 2350, 2351, See Table 2,
Morphogenic kinase assay. GSK3 kinase activity are Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; 22359, 2362, 2367, 2369, SEQ ID NO: Z
Protein 9 (BMP9); well known in the art. For Insulin resistance; Insulin deficiency; 22370, 2473, for particular
Growth example, Biol. Chem. 379(8-9): Hyperlipidemia; Hyperketonemia; Non-insulin 2623, 2624, construct.
differentiation (1998) 1101-1110.; dependent Diabetes Mellitus (NIDDM); Insulin- 2625, 2631,
factor-2 precursor Biochem J. 1993 Nov 15; 296 dependent Diabetes Mellitus (IDDM); A Condition 2632, 2633.
(GDF-2 (Pt 1): 15-9. Associated With Diabetes Including, But Not
precursor)) Limited To Obesity, Heart Disease, Hyperglycemia,
Infections, Retinopathy, And/Or Ulcers; Metabolic
Disorders; Immune Disorders; Obesity; Vascular
Disorders; Suppression of Body Weight;
Suppression of Appetite; Syndrome X.
Glucagon-Like- Stimulates the synthesis GLP1 activity may be assayed Hyperglycemia; Diabetes; Diabetes Insipidus; 2448, 2455, 2456, 2457, 2803, See Table 2, SEQ
Peptide 1 (GLP1; and release of insulin; in vitro using a [3-H]-glucose Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; 2804, 2900, 2904, 2945, 2964, ID NO: Z for
Insulinotropin) enhances the sensitivity of uptake assay. (J Biol Chem Insulin resistance; Insulin deficiency; Hyperlipidemia; 2982, 3070, 2802, 3027, 3028, particular
adipose, muscle, and liver 1999 Oct 22; 274(43): 30864-30873). Hyperketonemia; Non-insulin dependent Diabetes 3045, 3046, 3069, 3071, 3072, construct.
tissues towards insulin; Mellitus (NIDDM); Insulin-dependent Diabetes 3085, 3086, 3087, 3140, 3309
stimulates glucose uptake; Mellitus (IDDM); A Condition Associated With
slows the digestive process; Diabetes Including, But Not Limited To Obesity,
suppresses appetite; blocks Heart Disease, Hyperglycemia, Infections,
the secretion of glucagon. Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Exendin-4 (AC- Stimulates the synthesis Exendin-4 activity may be Hyperglycemia; Diabetes; Diabetes Insipidus; 2469 and 2470. See Table 2, SEQ
2993) and release of insulin; assayed in vitro using a [3-H]- Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; ID NO: Z for
enhances the sensitivity of glucose uptake assay. (J Biol Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
adipose, muscle, and liver Chem 1999 Oct 22; Hyperketonemia; Non-insulin dependent Diabetes construct.
tissues towards insulin; 274(43): 30864-30873). Mellitus (NIDDM); Insulin-dependent Diabetes
stimulates glucose uptake; Mellitus (IDDM); A Condition Associated With
slows the digestive process; Diabetes Including, But Not Limited To Obesity,
suppresses appetite; blocks Heart Disease, Hyperglycemia, Infections,
the secretion of glucagon. Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
T20 (T20 HIV a peptide from residues Virus inhibition assays as HIV; AIDS; SIV (simian immunodeficiency virus) 7777, 2672, 2673 See Table 2, SEQ
inhibitory peptide, 643-678 of the HIV gp41 described in Zhang et al., Sept. infection. ID NO: Z for
DP178; DP178 HIV transmembrane protein 26 2002, Sciencexpress particular
inhibitory peptide) ectodomain which binds to (www.sciencexpress.org). construct.
gp41 in its resting state and
prevents transformation to
the fusogenic state
T1249 (T1249 HIV a second generation HIV Virus inhibition assays as HIV; AIDS; SIV (simian immunodeficiency virus) 9999, 2667, 2670, 2946 See Table 2, SEQ
inhibitory peptide; fusion inbitor described in Zhang et al., Sept. infection ID NO: Z for
T1249 anti-HIV 26 2002, Sciencexpress particular
peptide) (www.sciencexpress.org). construct.
Interferon Hybrids, Confers a range of cellular Anti-viral assay: Rubinstein S, Viral infections; HIV Infections; Hepatitis; Chronic 2875, 2872, 2876, 2874, 2873. See Table 2, SEQ
specifically responses including Familletti PC, Pestka S. (1981) Hepatitis; Hepatitis B; Chronic Hepatitis B; Hepatitis ID NO: Z for
preferred: antiviral, antiproliferative, Convenient assay for C; Chronic Hepatitis C; Hepatitis D; Chronic Hepatitis particular
IFNalpha A/D hybrid antitumor and interferons. J. Virol. 37(2): 755-8; D; Human Papillomavirus; Herpes Simplex Virus construct.
(BgIII version) immunomodulatory Anti-proliferation assay: Infection; External Condylomata Acuminata; HIV;
IFNalpha A/D hybrid activities; stimulate Gao Y, et al (1999) Sensitivity HIV Infection; Oncology; Cancer; Solid Tumors;
(PvuII version) production of two of an epstein-barr virus- Melanoma; Malignant Melanoma; Renal Cancer (e.g.,
IFNalpha A/F hybrid enzymes: a protein kinase positive tumor line, Daudi, to Renal Cell Carcinoma); Lung Cancer (e.g,. Non-Small
IFNalpha A/B hybrid and an oligoadenylate alpha interferon correlates with Cell Lung Cancer or Small Cell Lung Cancer) Colon
IFNbeta 1/alpha D synthetase. Also, expression of a GC-rich viral Cancer; Breast Cancer; Liver Cancer; Prostate Cancer;
hybrid (IFNbeta- modulates MHC antigen transcript. Mol Cell Biol. Bladder Cancer; Gastric Cancer; Sarcoma; AIDS-
1/alpha-1 hybrid) expression, NK cell activity 19(11): 7305-13. Related Kaposi's Sarcoma; Lymphoma; T Cell
IFNalpha/beta hybrid and IFNg production and Lymphoma; Cutaneous T-Cell Lymphoma; Non-
IL12 production in Hodgkin's Lymphoma; Brain Cancer; Glioma;
monocytes. Glioblastoma Multiforme; Cervical Dysplasia;
Leukemia; Preleukemia; Bone Marrow Disorders;
Bone Disorders; Hairy Cell Leukemia; Chronic
Myelogeonus Leukemia; Hematological Malignancies;
Hematological Disorders; Multiple Myeloma;
Bacterial Infections; Chemoprotection;
Thrombocytopenia; Multiple Sclerosis; Pulmonary
Fibrosis; Age-Related Macular Degeneration; Macular
Degeneration; Crohn's Disease; Neurological
Disorders; Arthritis; Rheumatoid Arthritis; Ulcerative
Colitis; Osteoporosis, Osteopenia, Osteoclastogenesis;
Fibromyalgia; Sjogren's Syndrome; Chronic Fatigue
Syndrome; Fever; Hemmorhagic Fever; Viral
Hemmorhagic Fevers; Hyperglycemia; Diabetes;
Diabetes Insipidus; Diabetes mellitus; Type 1 diabetes;
Type 2 diabetes; Insulin resistance; Insulin deficiency;
Hyperlipidemia; Hyperketonemia; Non-insulin
dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM); A Condition
Associated With Diabetes Including, But Not Limited
To Obesity, Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
B-type natriuretic stimulates smooth muscle Inhibition of angiotensin can be Congestive heart failure; cardiac volume overload; 3119, 8888. See Table 2, SEQ
peptide (BNP, brain relaxation and vasodilation, determined using assays known cardiac decompensation; Cardiac Failure; Left ID NO: Z for
natriuretic peptide) natriuresis, and suppression in the art, for example using an Ventricular Dysfunction; Dyspnea particular
of renin-angiotensin and in vitro proliferation assay with construct.
endothelin. rat cardiac fibroblasts as
described in Naunyn
Schmiedebergs Arch
Pharmacol 1999
May; 359(5): 394-9.
Vasodilation can be measured
in animals by measuring the
myogenic responses of small
renal arteries in an isobaric
arteriograph system (see Am J
Physiol Regul Integr Comp
Physiol 2002
Aug; 283(2): R349-R355).
Natriuesis is determined by
measuring the amount of
sodium in the urine.
α-defensin, including Suppression of HIV Virus inhibition assays as HIV, AIDS; ARC. 3208, 3209, 3210. See Table 2, SEQ
alpha 1 defensin, replication; active against described in Zhang et al., Sept. ID NO: Z for
alpha 2 defensin, bacteria, fungi, and 26 2002, Sciencexpress particular
alpha 3 defensin enveloped viruses. (www.sciencexpress.org). construct.
(myeloid-related
defensin; DEFA1;
neutrophil-specific
defensin; CAF)
Phosphatonin Regulation of phosphate Blood phosphate levels can be Hyperphosphatemia; Hyperphosphatemia in chronic 3238. See Table 2, SEQ
(matrix extracellular metabolism. measured using methods renal failure; hypophosphatemia; Osteomalacia; ID NO: Z for
phosphoglycoprotein; known in the art such as the Rickets; X-linked dominant hypophosphatemic particular
MEPE) Hypophosphatemic Rat rickets/osteomalacia (XLH); autosomal dominant construct.
Bioassay. Zoolog Sci 1995 hypophosphatemic rickets/osteomalacia (ADHR);
Oct; 12(5): 607-10. tumor-induced rickets/osteomalacia (TIO).
P1pal-12 (pepducin, Regulation of protease- Platelet aggregation can be Protection against systemic platelet activation, 3274. See Table 2, SEQ
PAR1-based activated receptor (PAR) measured using methods thrombus, heart attack, stroke, and/or coagulation ID NO: Z for
pepducin) signal transduction and known in the art such as disorders. particular
thrombin-mediated described in Nature Medicine construct.
aggregation of human 2002 Oct; 8(10): 1161-1165.
platelets.
P4pal-10 (pepducin, Regulation of protease- Platelet aggregation can be Protection against systemic platelet activation, 3275. See Table 2, SEQ
PAR4-based activated receptor (PAR) measured using methods thrombus, heart attack, stroke, and/or coagulation ID NO: Z for
pepducin) signal transduction and known in the art such as disorders. particular
thrombin-mediated described in Nature Medicine construct.
aggregation of human 2002 Oct; 8(10): 1161-1165.
platelets.
HRDFD27 Involved in the T-cell proliferation can be Chemoprotection; Adjunct to Chemotherapy; 2361 See Table 2, SEQ
proliferation of T cells; measured using assays known Inflammatory disorders; Cancer; Leukemia; ID NO: Z for
Production of TNFgamma. in the art. For example, Myelocytic leukemia; Neutropenia, Primary particular
“Lymphocytes: a practical neutropenias (e.g.; Kostmann syndrome); Secondary construct.
approach” edited by: SL Rowland, neutropenia; Prevention of neutropenia; Prevention
AJ McMichael - and treatment of neutropenia in HIV-infected patients;
chapter 6, pages 138-160 Prevention and treatment of neutropenia associated
Oxford University Press with chemotherapy; Infections associated with
(2000): and “Current Protocols neutropenias; Myelopysplasia; Autoimmune disorders;
on CD-ROM” section 3.12 Psoriasis; Mobilization of hematopoietic progenitor
Proliferation Assays for T-cell cells; Wound Healing; Autoimmune Disease;
Function John Wiley & Soncs, Transplants; Bone marrow transplants; Acute
Inc. (1999). myelogeneous leukemia; Lymphoma, Non-Hodgkin's
lymphoma; Acute lymphoblastic leukemia; Hodgkin's
disease; Accelerated myeloid recovery; Glycogen
storage disease
HWHGZ51 (CD59; Stimulates an immune The ability to affect Skeletal diseases and disorders; Musculoskeletal 2407, 2408 See Table 2, SEQ
Metastasis-associated response and induces chondrocyte differentiation can diseases and disorders; Bone fractures and/or breaks; ID NO: Z for
GPI-adhered protein inflammation by inducing be measured using methods Osteoporosis (postmenopausal, senile, or idiopathic particular
homolog) mononuclear cell, known in the art, such as juvenile); Gout and/or pseudogout; Paget's disease; construct.
eosinophil and PMN described in Bone (1995) Sep; Osteoarthritis; Tumors and/or cancers of the bone
infiltration; Inhibits growth 17(3): 279-86. (osteochondromas, benign chondromas,
of breast cancer, ovarian chondroblastomas, chondromyxoid fibromas, osteoid
cancer, leukemia, and osteomas, giant cell tumors, multiple myelomas,
melanoma; Overexpressed osteosarcomas, fibrosarcomas, malignant fibrous
in colon, lung, breast and histiocytomas, chondrosarcomas, Ewing's tumors,
rectal tumors; Regulates and/or malignant lymphomas); Bone and joint
glucose and/or FFA update infections (osteomyelitits and/or infectious arthritis);
by adipocytes and skeletal Charcot's joints; Heel spurs; Sever's disease; Sport's
muscle; Induces injuries; Cancer; Solid Tumors; Melanoma; Malignant
redifferentiation of Melanoma; Renal Cancer (e.g., Renal Cell
chondrocytes Carcinoma); Lung Cancer (e.g,. Non-Small Cell Lung
Cancer or Small Cell Lung Cancer) Colon Cancer;
Breast Cancer; Liver Cancer; Prostate Cancer; Bladder
Cancer; Gastric Cancer; Sarcoma; AIDS-Related
Kaposi's Sarcoma; Lymphoma; T Cell Lymphoma;
Cutaneous T-Cell Lymphoma; Non-Hodgkin's
Lymphoma; Brain Cancer; Glioma; Glioblastoma
Multiforme; Cervical Dysplasia; Leukemia;
Preleukemia; Bone Marrow Disorders; Bone
Disorders; Hairy Cell Leukemia; Chronic
Myelogeonus Leukemia; Hematological Malignancies;
Hematological Disorders; Multiple Myeloma; Kidney
diseases and disorders; Shonlein-Henoch purpura,
Berger disease, celiac disease, dermatitis
herpetiformis, Chron disease; Diabetes; Diabetes
Insipidus; Diabetes mellitus; Type 1 diabetes; Type 2
diabetes; Insulin resistance; Insulin deficiency;
Hyperlipidemia; Hyperketonemia; Non-insulin
dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM); A Condition
Associated With Diabetes Including, But Not Limited
To Obesity, Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X; Kidney disorders;
Hyperinsulinemia; Hypoinsulinemia; Immunological
disorders (e.g. arthritis, asthma, immunodeficiency
diseases, AIDS, rheumatoid arthritis, granulomatous
disease, inflammatory bowl disease, sepsis, acne,
neutropenia, neutrophilia, psoriasis, hypersensitivities,
T-cell mediated cytotoxicity, host-versus-graft disease,
autoimmunity disorders, demyelination, systemic
lupus erythematosis, drug induced hemolytic anemia,
rheumatoid arthritis, Sjorgren's disease, scleroderma)
C17 (cytokine-like Inhibits glucose and/or Proliferation of kidney Kidney diseases and disorders; Shonlein-Henoch 2489, 2490 See Table 2, SEQ
protein C17) FFA uptake by adipocytes; mesangial cells can be assayed purpura, Berger disease, celiac disease, dermatitis ID NO: Z for
Induces proliferation of using techniques described in J. herpetiformis, Chron disease; Diabetes; Diabetes; particular
kidney mesangial cells; Investig. Med. (1998) Aug; Insipidus; Diabetes mellitus; Type 1 diabetes; Type 2 construct.
Regulation of cytokine 46(6): 297-302. diabetes; Insulin resistance; Insulin deficiency;
production and antigen Hyperlipidemia; Hyperketonemia; Non-insulin
presentation dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM); A Condition
Associated With Diabetes Including, But Not Limited
To Obesity, Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X; Kidney disorders;
Hyperinsulinemia; Hypoinsulinemia; Hematopoietic
disorders; Immunological diseases and disorders;
Developmental diseases and disorders; Hepatic
diseases and disorders; Cancer (particularly leukemia);
Immunological disorders (e.g. arthritis, asthma,
immunodeficiency diseases, AIDS, rheumatoid
arthritis, granulomatous disease, inflammatory bowl
disease, sepsis, acne, neutropenia, neutrophilia,
psoriasis, hypersensitivities, T-cell mediated
cytotoxicity, host-versus-graft disease, autoimmunity
disorders, demyelination, systemic lupus
erythematosis, drug induced hemolytic anemia,
rheumatoid arthritis, Sjorgren's disease, scleroderma)
HDPBQ71 Regulates production and Such assays that may be used Blood disorders and infection (e.g., viral infections, 2515, 2545 See Table 2, SEQ
secretion of IFNgamma; or routinely modified to test tuberculosis, infections associated with chronic ID NO: Z for
Activation of myeloid cells immunomodulatory activity of granulomatosus disease and malignant osteoporosis); particular
and/or hematopoietic cells polypeptides of the invention Autoimmune disease (e.g., rheumatoid arthritis, construct.
(including antibodies and systemic lupus erythematosis, multiple sclerosis);
agonists or antagonists of the Immunodeficiency, boosting a T cell-mediated
invention) include the assays immune response, and suppressing a T cell-mediated
disclosed in Miraglia et al., J immune response; Inflammation and inflammatory
Biomolecular Screening 4: 193-204 disorders; Idiopathic pulmonary fibrosis; Neoplastic
(1999); Rowland et al., diseases (e.g., leukemia, lymphoma, melanoma);
““Lymphocytes: a practical Neoplasms and cancers, such as, for example,
approach”” Chapter 6: 138-160 leukemia, lymphoma, melanoma, and prostate, breast,
(2000); Gonzalez et al., J Clin lung, colon, pancreatic, esophageal, stomach, brain,
Lab Anal 8(5): 225-233 (1995); liver and urinary cancer;. Benign dysproliferative
Billiau et al., Ann NY Acad Sci disorders and pre-neoplastic conditions, such as, for
856: 22-32 (1998); Boehm et example, hyperplasia, metaplasia, and/or dysplasia;
al., Annu Rev Immunol Anemia; Pancytopenia; Leukopenia;
15: 749-795 (1997), and Thrombocytopenia; Hodgkin's disease; Acute
Rheumatology (Oxford) lymphocytic anemia (ALL); Plasmacytomas; Multiple
38(3): 214-20 (1999) myeloma; Burkitt's lymphoma; Arthritis; AIDS;
Granulomatous disease; Inflammatory bowel disease;
Sepsis; Neutropenia; Neutrophilia; Psoriasis;
Suppression of immune reactions to transplanted
organs and tissues; Hemophilia; Hypercoagulation;
Diabetes mellitus; Endocarditis; Meningitis; Lyme
Disease; Asthma; Allergy
Oscar (osteoclast- Regulator of osteoclast Assay to detect osteoclast Skeletal diseases and disorders; Musculoskeletal 2571, 2749 See Table 2, SEQ
associated receptor differentiation; regulator of differentiation is described in J. diseases and disorders; Bone fractures and/or breaks; ID NO: Z for
isoform-3) innate and adaptive Exp. Med. (2002) Jan 21; Osteoporosis (postmenopausal, senile, or idiopathic particular
immune responses 195(2): 201-9. juvenile); Gout and/or pseudogout; Paget's disease; construct.
Osteoarthritis; Tumors and/or cancers of the bone
(osteochondromas, benign chondromas,
chondroblastomas, chondromyxoid fibromas, osteoid
osteomas, giant cell tumors, multiple myelomas,
osteosarcomas, fibrosarcomas, malignant fibrous
histiocytomas, chondrosarcomas, Ewing's tumors,
and/or malignant lymphomas); Bone and joint
infections (osteomyelitits and/or infectious arthritis);
Charcot's joints; Heel spurs; Sever's disease; Sport's
injuries
Tumstatin (T5, T7 or Inhibits angiogenesis; A tumor cell proliferation assay Cancer; Solid Tumors; Melanoma; Malignant 2647, 2648, 2649, 2650, 2943, See Table 2, SEQ
T8 peptide; Inhibits tumor growth; is described in J. Biol. Chem. Melanoma; Renal Cancer (e.g., Renal Cell 2944, 3047, 3048 ID NO: Z for
α3(IV)NC1) Inhibits protein synthesis (1997) 272: 20395-20401. Carcinoma); Lung Cancer (e.g,. Non-Small Cell Lung particular
Protein synthesis can be Cancer or Small Cell Lung Cancer) Colon Cancer; construct.
measured as described in Breast Cancer; Liver Cancer; Prostate Cancer; Bladder
Science (2002) Jan 4; Cancer; Gastric Cancer; Sarcoma; AIDS-Related
295(5552): 140-3. Kaposi's Sarcoma; Lymphoma; T Cell Lymphoma;
Cutaneous T-Cell Lymphoma; Non-Hodgkin's
Lymphoma; Brain Cancer; Glioma; Glioblastoma
Multiforme; Cervical Dysplasia; Leukemia;
Preleukemia; Bone Marrow Disorders; Bone
Disorders; Hairy Cell Leukemia; Chronic
Myelogeonus Leukemia; Hematological Malignancies;
Hematological Disorders; Multiple Myeloma;
Angiogenesis
CNTF (Ciliary Enhances myelin Regulation of myelin formation Neurological and neural diseases and disorders, 2724, 2725, 3171, 3172 See Table 2, SEQ
neurotrophic factor) formation; Reduces can be assayed as described in particularly diseases and disorders associated with ID NO: Z for
photoreceptor degredation; J. Neurosci. (2002) Nov. 1; myelin and demyelination, such as, for example, ALS, particular
Regulates calcium currents 22(21): 9221-7. multiple sclerosis, Huntington's disease; Neuronal and construct.
spinal cord injuries; Disorders of the eye, such as, for
example, retinitis pigmentosa, blindness, color-
blindness, macular degeneration.
Somatostatin Inhibits growth hormone, Inhibition of growth hormone Cancer; Metastatic carcinoid tumors; Vasoactive 2798, 2825, 2830, 2831, 2902 See Table 2, SEQ
(Octreotide; glucagons and insulin; release in humans by Intestinal Peptide secreting adenomas; Diarrhea and ID NO: Z for
octreotide acetate; Suppresses LF response to somatostatin can be measured Flushing; Prostatic disorders and cancers; Breast particular
Sandostating LAR ®) GnRH; Decreases as described in J. Clin. cancer; Gastrointestinal disorders and cancers; construct.
splanchnic blood flow; Endocrinol. Metab. (1973) Oct; Cancers of the endocrine system; Head and neck
Inhibits release of 37(4): 632-4. paragangliomas; Liver disorders and cancers;
serotonin, gastrin, Inhibition of insulin secretion Nasopharyngeal cancers; Thyroid disorders and
vasoactive intestinal by somatostatin can be cancers; Acromegaly; Carcinoid Syndrome;
peptide, secretin, motilin, measured as described in the Gallbladder disorders, such as gallbladder contractility
and pancreatic polypeptide. Lancet (1973) Dec. 8; diseases and abnormal bile secretion; Psoriasis;
2(7841): 1299-1301. Diabetes; Diabetes Insipidus; Diabetes mellitus; Type
1 diabetes; Type 2 diabetes; Insulin resistance; Insulin
deficiency; Hyperlipidemia; Hyperketonemia; Non-
insulin dependent Diabetes Mellitus (NIDDM);
Insulin-dependent Diabetes Mellitus (IDDM); A
Condition Associated With Diabetes Including, But
Not Limited To Obesity, Heart Disease,
Hyperglycemia, Infections, Retinopathy, And/Or
Ulcers; Metabolic Disorders; Immune Disorders;
Obesity; Vascular Disorders; Suppression of Body
Weight; Suppression of Appetite; Syndrome X;
Kidney disorders; Neurological disorders and diseases,
including Alzheimers Disease, Parkinson's disease and
dementia; Neuropsychotic disorders, including Bipolar
affective disorder; Rheumatoid arthritis; Hypertension;
Intracranial hypertension; Esophageal varices; Graves'
disease; Seizures; Epilepsy; Gastritis; Angiogenesis;
IL-22 (IL22, Stimulates glucose uptake IL-22 activity may be assayed Hyperglycemia; Diabetes; Diabetes Insipidus; 2901, 2903 See Table 2, SEQ
interleukin-22; in skeletal muscle cells; in vitro using a [3-H]-glucose Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; ID NO: Z for
IL17D, IL27) increases skeletal muscle uptake assay. (J Biol Chem Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
insulin sensitivity. 1999 Oct 22; 274(43): 30864-30873). Hyperketonemia; Non-insulin dependent Diabetes construct.
Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
HCE1P80 Stimulates glucose uptake HCE1P80 activity may be Hyperglycemia; Diabetes; Diabetes Insipidus; 2908, 3049, 3050, 3051, 3052 See Table 2, SEQ
in; increases insulin assayed in vitro using a [3-H]- Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; ID NO: Z for
sensitivity. glucose uptake assay. (J Biol Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
Chem 1999 Oct 22; Hyperketonemia; Non-insulin dependent Diabetes construct.
274(43): 30864-30873). Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
HDRMI82 Stimulates glucose uptake; HDRMI82 activity may be Hyperglycemia; Diabetes; Diabetes Insipidus; 2909 See Table 2, SEQ
increases insulin assayed in vitro using a [3-H]- Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; ID NO: Z for
sensitivity. glucose uptake assay. (J Biol Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
Chem 1999 Oct 22; Hyperketonemia; Non-insulin dependent Diabetes construct.
274(43): 30864-30873). Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
HDALV07 Modulates insulin action Insulin activity may be assayed Diabetes; Diabetes Insipidus; Diabetes mellitus; Type 3053, 3055, 3056 See Table 2, SEQ
(adiponectin; gelatin- in vitro using a [3-H]-glucose 1 diabetes; Type 2 diabetes; Insulin resistance; Insulin ID NO: Z for
binding 28k protein uptake assay. (J Biol Chem deficiency; Hyperlipidemia; Hyperketonemia; Non- particular
precurson; adipose 1999 Oct 22; 274(43): 30864-30873). insulin dependent Diabetes Mellitus (NIDDM); construct.
most abundant gene Insulin-dependent Diabetes Mellitus (IDDM); A
transcript; APM-1; Condition Associated With Diabetes Including, But
GBP28; ACRP30; Not Limited To Obesity, Heart Disease,
ADIPOQ) Hyperglycemia, Infections, Retinopathy, And/Or
Ulcers; Metabolic Disorders; Immune Disorders;
Obesity; Vascular Disorders; Suppression of Body
Weight; Suppression of Appetite; Syndrome X;
Hyperglycemia; Familial combined hyperlipidemia;
Metabolic syndrome; Inflammatory disorders;
Atherogenic disorders
C Peptide An insulin precursor C-peptide concentrations can Diabetes; Diabetes Insipidus; Diabetes mellitus; Type 3088, 3149 See Table 2, SEQ
involved in insulin be measured using assays well 1 diabetes; Type 2 diabetes; Insulin resistance; Insulin ID NO: Z for
regulation known in the art, such as the deficiency; Hyperlipidemia; Hyperketonemia; Non- particular
one described in PNAS (1970) insulin dependent Diabetes Mellitus (NIDDM); construct.
Sep; 67(1): 148-55 Insulin-dependent Diabetes Mellitus (IDDM); A
Condition Associated With Diabetes Including, But
Not Limited To Obesity, Heart Disease,
Hyperglycemia, Infections, Retinopathy, And/Or
Ulcers; Metabolic Disorders; Immune Disorders;
Obesity; Vascular Disorders; Suppression of Body
Weight; Suppression of Appetite; Syndrome X;
Hyperglycemia; Familial combined hyperlipidemia;
Metabolic syndrome
HCBOG68 (enteric Controls proliferation/ Activation of cAMP-mediated Treatment of Obesity; treatment of Diabetes; 3106, 3270 See Table 2, SEQ
adipokine; Fat SID; differentiation or transcription in adipocytes can suppression of body weight gain; suppression of ID NO: Z for
proline rich acidic metabolism/ be assayed using methods appetite. Hyperglycemia; Diabetes; Diabetes Insipidus; particular
protein) physiology/pathology/of known in the art (Berger et al., Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; construct.
adipocytes and adipose Gene 66: 1-10 (1998); Cullen Insulin resistance; Insulin deficiency; Hyperlipidemia;
tissue in response to dietary and Malm, Methods in Hyperketonemia; Non-insulin dependent Diabetes
conditions. Enzymol 216: 362-368 (1992); Mellitus (NIDDM); Insulin-dependent Diabetes
Henthorn et al., Proc Natl Acad Mellitus (IDDM); A Condition Associated With
Sci USA 85: 6342-6346 (1988); Diabetes Including, But Not Limited To Obesity,
Reusch et al., Mol Cell Biol Heart Disease, Hyperglycemia, Infections,
20(3): 1008-1020 (2000); and Retinopathy, And/Or Ulcers; Metabolic Disorders;
Klemm et al., J Biol Chem Immune Disorders; Obesity; Vascular Disorders;
273: 917-923 (1998)). Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Other indications for antibodies and/or antagonists,
include treatment of weight loss; treatment of AIDS
wasting; appetite stimulant; treatment of cachexia.
PYY (Peptide YY), Decreases appetite; Appetite and food intake can be Most preferred: Treatment of Obesity; treatment of 3108, 3109, 3281, 3117, 3118, See Table 2, SEQ
including PYY3-36 increases satiety; decreases can be measured by methods Diabetes; suppression of body weight gain; 3282. ID NO: Z for
(amino acid residues food intake. known in the art (Batterham et suppression of appetite. particular
31-64 of full length al. Nature 2002; 418: 650654) Hyperglycemia; Diabetes; Diabetes Insipidus; construct.
PYY, amino acid Diabetes mellitus; Type 1 diabetes; Type 2 diabetes;
residues 3-36 of Insulin resistance; Insulin deficiency; Hyperlipidemia;
mature PYY) Hyperketonemia; Non-insulin dependent Diabetes
Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Other indications for antibodies, antagonists: treatment
of weight loss; treatment of AIDS wasting; appetite
stimulant; treatment of cachexia.
WNT10b Inhibits adipogenesis. WNT10b activity can be Most preferred: Treatment of Obesity; suppression of 3141 See Table 2, SEQ
measured using adipogenesis body weight gain; suppression of appetite. ID NO: Z for
inhibition assays (Ross et al., Other indications: Hyperglycemia; Diabetes; Diabetes particular
Science 2000; 289(5481): 950-953 Insipidus; Diabetes mellitus; Type 1 diabetes; Type 2 construct.
diabetes; Insulin resistance; Insulin deficiency;
Hyperlipidemia; Hyperketonemia; Non-insulin
dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM).
WNT11 Promotes cardiogenesis. WNT11 activity can be Treatment of Cardiovascular disorders; Congestive 3142 See Table 2, SEQ
measured using assays known Heart Failure; Myocardial Infarction. ID NO: Z for
in the art, including particular
cardiogenesis assays (Eisenberg construct.
et al., Dev Dyn 1999
Sep; 216(1): 45-58).
Herstatin Inhibits cancer Herstatin activity can be Oncology; Cancer; Solid Tumors; Melanoma; 3143 See Table 2, SEQ
proliferation. measured using cell Malignant Melanoma; Renal Cancer (e.g., Renal Cell ID NO: Z for
proliferation assays known in Carcinoma); Lung Cancer (e.g,. Non-Small Cell Lung particular
the art (Doherty et al., PNAS Cancer or Small Cell Lung Cancer); Colon Cancer; construct.
1999; 96(19): 10869-10874. Breast Cancer; Liver Cancer; Prostate Cancer; Bladder
Cancer; Gastric Cancer; Sarcoma; AIDS-Related
Kaposi's Sarcoma; Lymphoma; T Cell Lymphoma;
Cutaneous T-Cell Lymphoma; Non-Hodgkin's
Lymphoma; Brain Cancer; Glioma; Glioblastoma
Multiforme; Cervical Dysplasia; Leukemia;
Preleukemia; Hairy Cell Leukemia; Chronic
Myelogeonus Leukemia; Hematological Malignancies;
Hematological Disorders; Multiple Myeloma.
Adrenomedullin stimulates vasodilation; Vasodilation can be measured Treatment of Congestive Heart Failure; Hypertension; 3144 See Table 2, SEQ
promotes bone growth. using assays known in the art Myocardial Infarction; Septic Shock; Osteoporosis; ID NO: Z for
(Ashton et al. Pharmacology Postmenopausal osteoporosis; Osteopenia. particular
2000; 61(2): 101-105. The construct.
promotion of bone growth can
be measured using assays
known in the art, such as the
osteoblast proliferation assay
(Cornish et al. Am J Physiol
1997 Dec; 273(6 Pt 1): E1113-20).
Nogo Receptor Receptor for the axon The promotion of axon Treatment of Central Nervous System Damage; Spinal 3184, 3185 See Table 2, SEQ
growth inhibitor, Nogo. regeneration and growth can be Cord Injury; Peripheral Nerve Damage; ID NO: Z for
measured using assays known Neurodegenerative Diseases; Parkinson's Disease; particular
in the art (Fournier et al. Nature Alzheimer's Disease; Huntington's Disease; construct.
2001; 409(6818): 341-346). Amyotrophic Lateral Sclerosis; Progressive
Supranuclear Palsy; Creutzfeld-Jacob Disease; Motor
Neuron Disease.
CART (Cocaine- and Inhibits food intact and fat Appetite and food intake can be Most preferred: Treatment of Obesity; suppression of 3232 See Table 2, SEQ
Amphetamine- storage; promotes lipid can be measured by methods body weight gain; suppression of appetite. ID NO: Z for
Regulated oxidation. known in the art (Batterham et Other indications: Hyperglycemia; Diabetes; Diabetes particular
Transcript) al. Nature 2002; 418: 650654) Insipidus; Diabetes mellitus; Type 1 diabetes; Type 2 construct.
diabetes; Insulin resistance; Insulin deficiency;
Hyperlipidemia; Hyperketonemia; Non-insulin
dependent Diabetes Mellitus (NIDDM); Insulin-
dependent Diabetes Mellitus (IDDM).
RegIV (Colon Stimulates glucose uptake; RegIV activity may be assayed Hyperglycemia; Diabetes; Diabetes Insipidus; 2910. See Table 2, SEQ
Specific Gene; Colon increases insulin in vitro using a [3-H]-glucose Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; ID NO: Z for
Specific Protein) sensitivity. uptake assay. (J Biol Chem Insulin resistance; Insulin deficiency; Hyperlipidemia; particular
1999 Oct 22; 274(43): 30864-30873). Hyperketonemia; Non-insulin dependent Diabetes construct.
Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Cosyntropin Synthetic corticotropin; The activity of cosyntropin can Endocrine; Addison's disease; Cushing's syndrome; SEQ ID: NO: 2198
(Cortrosyn) stimulates the release of be assessed in vivo by pituitary dysfunction; acute adrenal crisis
(CAS-16960-16-0) cortisol. measuring serum cortisol
levels. (Frank et al. J. Am. Vet.
Med. Assoc. 1998
212(10): 1569-71).
Pexiganan Acetate Disrupts bacterial Pexiganan acetate activity can Treatment of Infectious Diseases; Treatment of SEQ ID NO:
(CAS-172820-23-4) membranes. be assessed using in vitro Bacterial Infections. 2199
antibacterial assays known in
the art. (Zasloff et al.,
Antimicrobial Agents and
Chemotherapy 1999, 43: 782-788).
Pramlintide (Amylin) Slows gastric emptying; Appetite and food intake can be Treatment of Obesity; treatment of Diabetes; SEQ ID NO:
(CAS-151126-32-8) decreases food intake. can be measured by methods suppression of body weight gain; suppression of 2200
known in the art (Batterham et appetite; treatment of endocrine disorders;
al. Nature 2002; 418: 650654) Hyperglycemia; Diabetes; Diabetes Insipidus;
Diabetes mellitus; Type 1 diabetes; Type 2 diabetes;
Insulin resistance; Insulin deficiency; Hyperlipidemia;
Hyperketonemia; Non-insulin dependent Diabetes
Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Other indications for antibodies, antagonists: treatment
of weight loss; treatment of AIDS wasting; appetite
stimulant; treatment of cachexia.
Teriparatide Acts in conjuction with Adenylyl cyclase stimulation in Bone Disorders; Fracture prevention; Hypercalcemia; SEQ ID NO:
(CAS-52232-67-4) calcitonin to control rat osteosarcoma cells, Malignant hypercalcemia; Osteoporosis; Paget's 2201
calcium and phosphate ovariectomized rat model of disease; Osteopenia, Osteoclastogenesis; osteolysis;
metabolism; elevates blood osteoporosis: IUBMB Life osteomyelitis; osteonecrosis; periodontal bone loss;
calcium level; stimulates 2000 Feb; 49(2): 131-5 osteoarthritis; rheumatoid arthritis; osteopetrosis;
the activity of osteocytes; periodontal, lytic, or metastatic bone disease;
enhances absorption of osteoclast differentiation inhibition; bone disorders;
Ca+/Pi from small intestine bone healing and regeneration.
into blood; promotes
reabsorption of Ca+ and
inhibits Pi by kidney
tubules.
Terlipressin Analog of vasopressin; Terlipressin activity can be Variceal hemorrhage; cirrhosis; portal hypertension; SEQ ID NO:
(triglycyl lycine induces vasoconstriction. measured using assays of hepatorenal syndrome; Blood-related disorders 2202
vasopressin) vasoconstriction, such as the
(CAS-14636-12-5) isolated arterial ring
preparation. (Landstrom et al.,
Hum Reprod 1999
Jan; 14(1): 151-5).
Ularitide Stimulates natriuresis, Ularitide activity can be Excretory disorders; Acute renal failure; asthma; SEQ ID NO:
(CAS-118812-69-4) diuresis, and vasodilation. assessed by measuring cGMP congestive heart failure; hypertension; pulmonary 2203
accumulation in rat renal cells. hypertension; cardiovascular disorders
(Valentin et al., Hypertension
1993 Apr; 21(4): 432-8).
Aprotinin (Trasylol) Serine protease inhibitor; Inhibition of thrombin-induced Inhibition of fibrinolysis; reduction of blood loss SEQ ID NO:
(CAS-9087-70-1; attenuates Systemic platelet aggregation can be during surgery; Treatment of Inflammation and 2204
CAS-11061-94-2; Inflammatory Response, measured using methods Immune Disorders.
CAS-12407-79-3) fibrinolysis and thrombin- known in the art. (Poullis et al.,
induced platelet J Thorac Cardiovasc Surg 2000
aggregation. Aug; 120(2): 370-8).
Aspartocin Antibacteria Aspartocin activity can be Treatment of Infectious Diseases; treatment of SEQ ID NO:
(CAS-4117-65-1; assessed using in vitro bacterial infections. 2205
CAS-1402-89-7) antibacterial assays known in
the art. (Zasloff et al.,
Antimicrobial Agents and
Chemotherapy 1999, 43: 782-788).
Calcitonin Regulates levels of calcium Hypocalcemic Rat Bioassay, Musculoskeletal; Osteroporosis; Paget's disease; SEQ ID NO:
(Calcimar) and phosphate in serum; bone resorbing assay and the hypercalcemia; 2206
(CAS-21215-62-3) causes a reduction in serum pit assay, CT receptor binding Bone Disorders; Fracture prevention; Malignant
calcium--an effect opposite assay, CAMP stimulation hypercalcemia; Osteopenia, Osteoclastogenesis;
to that of human assay: J Bone Miner Res 1999 osteolysis; osteomyelitis; osteonecrosis; periodontal
parathyroid hormone. Aug; 14(8): 1425-31 bone loss; osteoarthritis; rheumatoid arthritis;
osteopetrosis; periodontal, lytic, or metastatic bone
disease; osteoclast differentiation inhibition; bone
disorders; bone healing and regeneration.
Carperitide (HANP; Stimulates natriuresis, Carperitide activity can be Treatment of Heart Failure; Cardiovascular disorders; SEQ ID NO:
recombinant human diuresis, and vasodilation. assessed in vitro by measuring Respiratory disorders; Acute respiratory distress 2207
atrial natriuretic cGMP accumulation in a syndrome.
peptide) number of cell lines, including
(CAS-89213-87-6) PC12 cells and cultured human
glomerular cells. (Medvede et
al., Life Sci 2001 Aug
31; 69(15): 1783-90; Green et
al., J Am Soc Nephrol 1994
Oct; 5(4): 1091-8).
Desirudin Inhibits thrombin; inhibits Desirudin activity can be Blood-related disorder; Thrombosis; SEQ ID NO:
(recombinant blood clotting. assessed using blood clotting thrombocytopenia; hemorrhages. 2208
hirudin; Revasc) assays known in the art, such as
(CAS-120993-53-5) in vitro platelet aggragation
assays. (Glusa, Haemostasis
1991; 21 Suppl 1: 116-20).
Emoctakin proinflammatory cytokine Treatment of Inflammation, Immune disorders, RSV SEQ ID NO:
(interleukin 8) infection. 2209
(CAS-142298-00-8)
Felypressin Derivative of Vasopressin; Felypressin vasoconstriction Treatment of pain; to induce local anesthesia. SEQ ID NO:
(CAS-56-59-7) Stimulates activity can be measured using 2210
vasoconstriction; Induces assays of vasoconstriction, such
local anesthesia. as the isolated arterial ring
preparation. (Landstrom et al.,
Hum Reprod 1999
Jan; 14(1): 151-5).
Glucagon Induces hyperglycemia. Glucagon activity may be Hypoglycemia; Diabetes; Diabetes Insipidus; Diabetes SEQ ID NO:
(CAS-16941-32-5) assayed in vitro using a [3-H]- mellitus; Type 1 diabetes; Type 2 diabetes; Insulin 2211
glucose uptake assay. (J Biol resistance; Insulin deficiency; Hyperlipidemia;
Chem 1999 Oct 22; Hyperketonemia; Non-insulin dependent Diabetes
274(43): 30864-30873). Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X; Endocrine disorders.
Nagrestipen Inflammation; Immune SEQ ID NO:
(CAS-166089-33-4) 2212
Pentigetide (Pentyde) Respiratory; Allergy; Immune SEQ ID NO:
(CAS-62087-72-3) 2213
Proinsulin Stimulates glucose uptake Insulin activity may be assayed Hyperglycemia; Diabetes; Diabetes Insipidus; SEQ ID NO:
(CAS-67422-14-4) and promotes glycogenesis in vitro using a [3-H]-glucose Diabetes mellitus; Type 1 diabetes; Type 2 diabetes; 2214
and lipogenesis. uptake assay. (J Biol Chem Insulin resistance; Insulin deficiency; Hyperlipidemia;
1999 Oct 22; 274(43): 30864-30873). Hyperketonemia; Non-insulin dependent Diabetes
Mellitus (NIDDM); Insulin-dependent Diabetes
Mellitus (IDDM); A Condition Associated With
Diabetes Including, But Not Limited To Obesity,
Heart Disease, Hyperglycemia, Infections,
Retinopathy, And/Or Ulcers; Metabolic Disorders;
Immune Disorders; Obesity; Vascular Disorders;
Suppression of Body Weight; Suppression of
Appetite; Syndrome X.
Becaplermin Promotes wound healing. Becaplermin activity can be Stimulate Epithelial Cell Proliferation; Stimulate Basal SEQ ID NO:
(Regranex; assessed using animal wound Keratinocytes; Promote Wound Healing; Stimulate 2215
recombinant PDGF- healing models known in the Hair Follicle Production; Healing Of Dermal Wounds.
BB) art. (Saba et al., Ann Plast Surg Wound Healing; Eye Tissue Wounds, Dental Tissue
(CAS-165101-51-9) 2002 Jul; 49(1): 62-6). Wounds, Oral Cavity Wounds, Diabetic Ulcers,
Dermal Ulcers, Cubitus Ulcers, Arterial Ulcers,
Venous Stasis Ulcers, Burns Resulting From Heat
Exposure Or Chemicals, or Other Abnormal Wound
Healing Conditions such as Uremia, Malnutrition,
Vitamin Deficiencies or Complications Associated
With Systemic Treatment With Steroids, Radiation
Therapy or Antineoplastic Drugs or Antimetabolites;
Promote Dermal Reestablishment Subsequent To
Dermal Loss; Increase the Adherence Of Skin Grafts
To A Wound Bed; Stimulate Re-Epithelialization from
The Wound Bed; To Promote Skin Strength; Improve
The Appearance Of Aged Skin; Proliferate
Hepatocytes, Lung, Breast, Pancreas, Stomach,
Bladder, Small Intestine, Large Intestine; Sebocytes,
Hair Follicles, Type II Pneumocytes, Mucin-
Producing Goblet Cells, or Other Epithelial Cells,
Endothelial Cells, Keratinocytes, or Basal
Keratinocytes (and Their Progenitors) Contained
Within The Skin, Lung, Liver, Bladder, Eye, Salivary
Glands, or Gastrointestinal Tract; Reduce The Side
Effects Of Gut Toxicity That Result From Radiation,
Chemotherapy Treatments Or Viral Infections;
Cytoprotector, especially of the Small Intestine
Mucosa or Bladder; Mucositis (Mouth Ulcers);
Regeneration Of Skin; Full and/or Partial Thickness
Skin Defects, including Burns, (e.g., Repopulation Of
Hair Follicles, Sweat Glands, And Sebaceous Glands);
Psoriasis; Epidermolysis Bullosa; Blisters; Gastric
and/or Doudenal Ulcers; Reduce Scarring;
Inflamamatory Bowel Diseases; Crohn's Disease;
Ulcerative Colitis; Gut Toxicity; Lung Damage;
Repair Of Alveoli And/or Brochiolar Epithelium;
Acute Or Chronic Lung Damage; Emphysema, ARDS;
Inhalation Injuries; Hyaline Membrane Diseases;
Infant Respiratory Distress Syndrome;
Bronchopulmonary Displasia In Premature Infants;
Fulminant Liver Failure; Cirrhosis, Liver Damage
caused by Viral Hepatitis and/or Toxic Substances;
Diabetes Mellitus; Inflammation; Cancer; Digestive
disorders.
Ghrelin Stimulates release of Appetite and food intake can be Endocrine; loss of body weight; loss of body weight SEQ ID NO:
(Genbank Accession growth hormone from can be measured by methods associated with cancer or anorexia nervosa; loss of 2216
No. AB029434) anterior pituitary. known in the art (Batterham et appetite; excessive appetite; body weight gain;
Stimulates appetite and al. Nature 2002; 418: 650654) Obesity; Diabetes; Acromegaly; Growth failure;
reduces fat burning. Growth hormone deficiency; Growth failure and
growth retardation Prader-Willi syndrome in children
2 years or older; Growth deficiencies; Growth failure
associated with chronic renal insufficiency;
Postmenopausal osteoporosis; burns; cachexia; cancer
cachexia; dwarfism; metabolic disorders; obesity;
renal failure; Turner's Syndrome, pediatric and adult;
fibromyalgia; fracture treatment; frailty, AIDS wasting
Ghrelin-binding Inhibits growth hormone Appetite and food intake can be Endocrine; Obesity; Diabetes; body weight gain;
antibody including release in response to can be measured by methods excessive appetite; loss of appetite; loss of body
antibody fragment, Ghrelin; inhibits increase in known in the art (Batterham et weight.
or dominant-negative appetite. al. Nature 2002; 418: 650654)
form of Ghrelin
receptor
NOGO-66 peptide Neurodegenerative disorders; spinal cord injury; SEQ ID NO:
fragment neuronal injury; brain trauma; stroke; multiple 2217
(Genbank Accession sclerosis; demyelinating disorders; neural activity and
No. NP_008939 neurological diseases; neural cell (e.g., neuron, glial
(amino acids 62-101)) cell, and schwann cell) regeneration and/or growth
Gastric inhibitory Increases nutrient uptake Nutrient uptake and Most preferred: loss of body weight, AIDS wasting, SEQ ID NO:
polypeptide (GIP), and tryglyceride tryglyceride accumulation can cachexia, loss of apetite. Other: Obesity; Diabetes; 2218
including GIP accumulation in be measured by methods insulin resistance; body weight gain; excessive
fragments adipocytes, which leads to desribed in Miyawaki et al., appetite.
(Genbank Accession obesity and insulin Nat. Medicine, 2002, Vol
No. NM_004123) resistance. 8(7): 738-742.
Gastric inhibitory Increased use of fat as Fat utilization as an energy Obesity; Diabetes; Insulin resistance; body weight
polypeptide predominant energy source; source can be measured as gain.
antibody, or antibody decreased accumulation of described in Miyawaki et al.,
fragments fat in adipocytes. Nat. Medicine, 2002, Vol
8(7): 738-742.
Gastric inhibitory Increased use of fat as Fat utilization as an energy Most preferred: Obesity; Diabetes; body weight gain; SEQ ID NO:
peptide receptor or predominant energy source; source can be measured as excessive appetite; insulin resistance. Other: loss of 2219
receptor fragments or decreased accumulation of described in Miyawaki et al., body weight, AIDS wasting, loss of appetite.
variants including fat in adipocytes. Nat. Medicine, 2002, Vol
soluble fragments or 8(7): 738-742.
variants
(Genbank Accession
Number
NM_000164)
POMC Activity of POMC-derived Preferred: resistance to stress; anti-inflammatory SEQ ID NO:
(proopiomelanocortin), fragments are diverse, and activity; analgesic activity; increased skin 2220
including well-known in the art. pigmentation; increased protein catabolism; increased
fragments or variants See, for example, Hadley et gluconeogenesis; obesity; diabetes. Other: decreased
(such as, for al., Ann N Y Acad Sci protein catabolism, decreased skin pigmentation,
example, alpha- 1999 Oct 20; 885: 1-21; Addison's disease, Cushing's syndrome
melanocyte Dores, Prog Clin Biol Res
stimulating hormone, 1990; 342: 22-7; Blalock,
αMSH, gamma Ann NY Acad Sci. 1999
melanocyte Oct 20; 885: 161-72).
stimulating hormone,
γMSH, beta-
melanocyte
stimulating hormone,
βMSH,
adrenocorticotropin,
ACTH, beta-
endorphin, met-
enkephalin)
(Genbank Accession
No. NM_000930)
HP 467, HP228 See US Patent No. See U.S. Pat. No. 6,350,430 Resistance to stress; anti-inflammatory activity; SEQ ID NO:
(U.S. Pat. No. 6,350,430 analgesic activity; increased skin pigmentation; 2221
6,350,430) increased protein catabolism; increased
gluconeogenesis.
NDP See U.S. Pat. No. See U.S. Pat. No. 6,350,430 Resistance to stress; anti-inflammatory activity; SEQ ID NO:
(U.S. Pat. No. 6,350,430 analgesic activity; increased skin pigmentation; 2222
6,350,430) increased protein catabolism; increased
gluconeogenesis.
Interleukin-21 Immunomodulator; inhibits IL-21 activity can be assessed Autoimmune disorders; Inflammatory disorders; 3298 SEQ ID NO:
(IL-21) interferon gamma by measuring interferon gamma Treatment of Psoriasis; Rheumatoid Arthritis; 2177
production by Th1 cells. production in Th1 cells. Inflammatory bowel disease.
(Wurster et al.,: J Exp Med
2002 Oct 7; 196(7): 969-77)
Interleukin-4 Immunomodulator; IL-4 activity can be assessed by Treatment of Psoriasis; Autoimmune disorders; 3307 SEQ ID NO:
(IL-4) promotes the differentiation measuring Th1/Th2 cytokine Rheumatoid Arthritis; Inflammatory bowel disease; 2178
of T cells into Th2 responses of isolated spleen Inflammatory disorders.
phenotype. cells in vitro. (Waltz et al.,
Horm Metab Res 2002
Oct; 34(10): 561-9).
Osteoclast Inhibitory Inhibits osteoclast Osteoclast Inhibitory Lectin Treatment of Bone Disorders; Osteoporosis; Fracture 3312 SEQ ID NO: 2181
Lectin formation. activity can be assessed using prevention; Hypercalcemia; Malignant hypercalcemia;
(OCIL) osteoclast formation assays Paget's disease; Osteopenia, Osteoclastogenesis;
known in the art. (Zhou et al., J osteolysis; osteomyelitis; osteonecrosis; periodontal
Biol Chem 2002 Dec bone loss; osteoarthritis; rheumatoid arthritis;
13; 277(50): 48808-15) osteopetrosis; periodontal, lytic, or metastatic bone
disease; osteoclast differentiation inhibition; bone
healing and regeneration.

TABLE 2
SEQ SEQ
Fusion Expression SEQ ID SEQ ID SEQ ID Leader
No. Construct ID Construct Name Description Vector NO: Y ID NO: X NO: Z ID NO: A NO: B Sequence
1 1520 pC4:HSA/TR6.V30-H300 Amino acids V30 to H300 of TR6 (fragment pC4 217 1 433 649 650 HSA
shown as V1 to H271 of SEQ ID NO: 433) fused
downstream of HSA.
2 1537 pYPG:HSA.TR6coV30-E294 Amino acids V30 to E294 of TR6 (fragment shown pYPGaf 218 2 434 651 652 HSA
as V1 to E265 of SEQ ID NO: 434) fused
downstream of HSA. DNA encoding TR6 has been
codon optimized.
3 1545 pYPG:HSA.TR6coV30-L288 Amino acids V30 to L288 of TR6 (fragment shown pYPGaf 219 3 435 653 654 HSA
as V1 to L259 of SEQ ID NO: 435) fused
downstream of HSA. DNA encoding TR6 has been
codon optimized.
4 1546 pYPG:HSA.TR6coV30-R284 Amino acids V30 to R284 of TR6 (fragment shown pYPGaf 220 4 436 655 656 HSA
as V1 to R255 of SEQ ID NO: 436) fused
downstream of HSA. DNA encoding TR6 has been
codon optimized.
5 1568 pSAC35:HSA-yTR6 TR6 fused downstream of HSA. DNA encoding pSAC35 221 5 437 657 658 HSA/kex2
TR6 has been codon optimized.
6 1570 pSAC35:TR6-HSA Mature TR6 fused downstream of the HSA/kex2 pSAC35 222 6 438 659 660 HSA/kex2
leader and upstream of the mature HSA.
7 1622 pC4:synTR6.M1-H300.HSA Synthetic TR6 fused upstream of mature HSA, pC4 223 7 439 661 662 Native TR6
with 2 extra amino acids between the TR6 and
HSA portions.
8 1623 pC4:HSA.synTR6.V30-H300 Synthetic mature TR6 fused downstream of FL pC4 224 8 440 663 664 HSA
HSA. Last amino acid HSA sequence is missing at
BSU36I site.
9 1642 pSAC35:GCSF.T31-P204. Mature GCSF cloned downstream of the HSA/kex2 pSAC35 225 9 441 665 666 HSA/kex2
HSA leader and upstream of the mature HSA
10 1643 pSAC35:HSA.GCSF.T31-P204 Mature GCSF cloned downstream of the mature pSAC35 226 10 442 667 668 HSA/kex2
HSA and HSA/kex2 leader sequence.
11 1645 pSAC35:yTR6(N173Q).HSA Mutant mature TR6 cloned upstream of mature pSAC35 227 11 443 669 670 HSA/kex2
HSA and downstream of the HSA/kex2 leader
sequence.
12 1657 pC4.HSA:DeCAF.A23-D233 Amino acids A23 to D233 of DeCAF fused pC4 228 12 444 671 672 HSA
downstream of full length HSA.
13 1680 pYPG:HSA.BLyS.A134-L285 Amino acids A134 to L285 of BLyS fused pYPGaf 229 13 445 673 674 HSA
downstream of FL HSA. Two extra amino acids
(Leu, Glu) have been added between the
therapeutic protein and HSA portions.
14 1681 pYPG.HSA.MPIF.D45-N120 Amino acids D45 to N120 of MPIF fused pYPGaf 230 14 446 675 676 HSA
downstream of FL HSA. Two additional amino
acids (L and E) have been added between HSA and
MPIF.
15 1697 pSAC35:HSA.GM-CSF.A18-E144 Amino acids A18 to E144 of GM-CSF fused pSAC35 231 15 447 677 678 HSA
downstream of FL HSA.
16 1699 pSAC35:GM-CSF.A18-E144: Amino acids A18 to E144 of GM-CSF fused pSAC35 232 16 448 679 680 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader.
17 1700 pSAC35:HSA-yTR6(N173Q) Mutant TR6 fused downstream of mature HSA pSAC35 233 17 449 681 682 HSA/kex2
with HSA/kex2 leader sequence.
18 1702 pYPG:HSA.ek.TR6coV30-L288 Amino acids V30 to L288 of TR6 (fragment shown pYPGaf 234 18 450 683 684 HSA
as V1 to L259 of SEQ ID NO: 450) fused
downstream of FL HSA with an enterokinase site
in between. DNA encoding TR6 has been codon
optimized.
19 1703 pYPG:HSA.ek.TR6coV30-R284 Amino acids V30 to R284 of TR6 (fragment shown pYPGaf 235 19 451 685 686 HSA
as V1 to R255 of SEQ ID NO: 451) fused
downstream of HSA with an enterokinase site in
between. DNA encoding TR6 has been codon
optimized.
20 1704 pYPG:HSA.TR6.V30-E294 Amino acids V30 to E294 of TR6 fused pYPGaf 236 20 452 687 688 HSA
downstream of HSA. Two additional amino acids
(Leu, Glu) are in between HSA and TR6.
21 1746 pYPG:HSA.ek.KDI.L28-K207 Amino acids L28 to K207 of KDI fused pYPGaf 237 21 453 689 690 HSA
downstream of HSA with an enterokinase site in
between.
22 1747 pSAC35.HSA.hGHRF.Y32-L75 Amino acids Y32 to L75 of hGHRF fused pSAC35 238 22 454 691 692 HSA
downstream of HSA.
23 1748 pSAC35.hGHRF.Y32-L75. Amino acids Y32 to L75 of hGHRF (see also SEQ pSAC35 239 23 455 693 694 HSA/kex2
HSA IDNO: 454) fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
24 1749 pSAC35:HSA.PTH.S1-F3 FL HSA fused upstream of amino acids S1-F34 of pSAC35 240 24 456 695 696 HSA
PTH
25 1750 pSAC35:PTH.S1-F34.HSA Amino acids 1-34 of PTH fused upstream of pSAC35 241 25 457 697 698 HSA/kex2
mature HSA and downstream of HSA/kex2 leader
sequence.
26 1757 pSAC35:IL2.A21-T153. Mature human IL-2 with a single amino acid pSAC35 242 26 458 699 700 HSA/kex2
145C/S.HSA mutation (C to S at position 145) cloned
downstream of the HSA/KEX2 leader and
upstream of mature HSA
27 1758 pSAC35:HSA.IL2.A21-T153. Mature human IL-2 with a single amino acid pSAC35 243 27 459 701 702 HSA/kex2
145C/S mutation (C to S at position 145) cloned
downstream of HSA with HSA/kex2 leader
sequence.
28 1772 pSAC:EPOco.A28-D192. Amino acids A28-D192 of EPO variant (where pSAC35 244 28 460 703 704 HSA/kex2
HSA glycine at amino acid 140 has been replaced with
an arginine) fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence. DNA
encoding EPO has been codon optimized.
29 1774 pSAC:HSA.EPOco.A28-D192. Amino acids A28-D192 of EPO variant (where pSAC35 245 29 461 705 706 HSA/kex2
glycine at amino acid 140 has been replaced with
an arginine) fused downstream of HSA with
HSA/kex2 leader sequence. DNA encoding EPO
has been codon optimized.
30 1777 pSAC35:TNFR2.L23-D257. Mature TNFR2 fused downstream of the pSAC35 246 30 462 707 708 HSA/kex2
HSA HSA/kex2 signal and upstream of mature HSA.
31 1778 pSAC35:IFNβ.M22-N187: Residues M22-N187 of full-length IFNb (shown as pSAC35 247 31 463 709 710 HSA/kex2
HSA M1 to N166 of SEQ ID NO: 463) fused upstream of
mature HSA and downstream of HSA/kex2 leader
sequence.
32 1779 pSAC35:HSA:IFNβ.M22-N187 Residues M22-N187 of full-length IFNb (shown as pSAC35 248 32 464 HSA/kex2
M1 to N166 of SEQ ID NO: 464) fused
downstream of HSA with HSA/kex2 leader
sequence.
33 1781 pSAC:EPOcoA28-D192.HSA Amino acids A28-D192 of EPO variant (where pSAC35 249 33 465 711 712 HSA/kex2
51N/S, 65N/S, 110N/s glycine at amino acid 140 has been replaced with
an arginine) fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
Glycosylation sites at amino acid 51, 65, 110 are
mutated from N to S residue. DNA encoding EPO
has been codon optimized.
34 1783 pSAC:HSA.EPOcoA28-D192. Amino acids A28-D192 of EPO variant (where pSAC35 250 34 466 713 714 HSA/kex2
51N/S, 65N/S, 110N/s glycine at amino acid 140 has been replaced with
an arginine) fused downstream of HSA with
HSA/kex2 leader sequence. Glycosylation sites at
amino acids 51, 65, 110 are mutated from N to S
residue. DNA encoding EPO has been codon
optimized.
35 1784 pSAC35:HSA.TNFR2.L23-D257 Mature TNFR2 fused downstream of FL HSA. pSAC35 251 35 467 715 716 HSA
36 1785 pSAC35:KGF2Δ28.A63-S208: Amino acids A63 to S208 of KGF2 fused upstream pSAC35 252 36 468 717 718 HSA/kex2
HSA of mature HSA and downstream of the HSA/kex2
signal peptide.
37 1786 pSAC35:HSA.KGF2{D}28.A63-S208 Amino acids A63 to S208 of KGF2 fused pSAC35 253 37 469 719 720 HSA
downstream of HSA.
38 1788 pSAC35:HSA.TR2.P37-A192 Amino acids P37 to A192 of TR2 fused pSAC35 254 38 470 721 722 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence.
39 1793 pSAC35:HSA.EPO.A28-D192 Amino acids A28-D192 of EPO variant (where pSAC35 255 39 471 HSA/kex2
(N51A, N65A, N110A) glycine at amino acid 140 has been replaced with
an arginine; see, for example, SEQ ID NO: 499)
fused downstream of HSA with HSA/kex2 leader
sequence. Glycosylation sites at amino acids 51,
65, 110 are mutated from N to A residue.
40 1794 pSAC35:HSA.EPO.A28-D192 Amino acids A28-D192 of the EPO variant (where pSAC35 256 40 472 HSA/kex2
glycine at amino acid 140 has been replaced with
an arginine; see, for example, SEQ ID NO: 499)
fused downstream of HSA with HSA/kex2 leader
sequence.
41 1809 pSAC35.MDC.G25-Q93. Amino acids P26 to Q93 of MDC with an N- pSAC35 257 41 473 723 724 HSA/kex2
HSA terminal methionine, fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
42 1812 pSAC35:IL2.A21-T153.HSA Amino acids A21 to T153 of IL-2 fused pSAC35 258 42 474 725 726 HSA/kex2
downstream of the HSA/kex2 leader and upstream
of mature HSA.
43 1813 pSAC35:HSA.IL2.A21-T153 Amino acids A21 to T153 of IL-2 fused pSAC35 259 43 475 727 728 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence.
44 1821 pSAC35:scFv116A01.HSA BLyS antibody fused upstream of mature HSA pSAC35 260 44 476 729 730 Modified
which lacks the first 8 amino acids and downstream HSA/kex2,
from the HSA/kex2 signal sequence which lacks lacking the
the last two amino acids. last two
amino acids
45 1830 pSAC35:HSA.KEX2.HAGDG59. Amino acids L19-Q300 of HAGDG59 fused pSAC35 261 45 477 731 732 HSA/kex2
L19-Q300 downstream of the HSA/kex2 signal, mature HSA
and KEX2 cleavage site.
46 1831 pSAC35:HAGDG59.L19-Q300. HSA/kex2 signal peptide followed by amino acids pSAC35 262 46 478 733 734 HSA/kex2
HSA L19-Q300 of HAGDG59 followed by mature HSA.
47 1833 pSAC35:humancalcitonin.C1-G33: Human Calcitonin (amino acids C98-G130 of SEQ pSAC35 263 47 479 735 736 HSA/kex2
HSA ID NO: 479) fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
48 1834 pSAC35:HSA.humancalcitonin. Human Calcitonin (amino acids C98-G130 of SEQ pSAC35 264 48 480 737 738 HSA
C1-G33 ID NO: 480) fused downstream of FL HSA.
49 1835 pSAC35:salmoncalcitonin.C1-G33: Salmon Calcitonin amino acids C1-G33 fused pSAC35 265 49 481 739 740 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
50 1836 pSAC35:HSA.salmoncalcitonin. Salmon Calcitonin amino acids C1-G33 fused pSAC35 266 50 482 741 742 HSA
C1-G33 downstream of HSA.
51 1853 pSAC35:PTH(1-34)N26.HSA Amino acids 1 to 34 of PTH fused upstream of pSAC35 267 51 483 743 744 HSA/kex2
mature HSA and downstream of HSA/kex2 leader
sequence. Amino acid K26 of PTH mutated to
N26.
52 1854 pSAC35:HSA.PTH(1-34)N26 Amino acids 1 to 34 of PTH fused downstream of pSAC35 268 52 484 745 746 HSA
HSA. Amino acid K26 of PTH mutated to N26.
53 1862 pSAC35:HSA.GnRH.Q24-G33 Amino acids Q24-G33 of human gonadotropin pSAC35 269 53 485 747 748 HSA/kex2
releasing hormone fused downstream of HSA with
HSA/kex2 leader sequence.
54 1863 pSAC35:GnRHQ24-G33. Amino acids Q24-G33 of human gonadotropin pSAC35 270 54 486 749 750 HSA/kex2
HSA releasing hormone fused upstream of mature HSA
and downstream of HSA/kex2 leader sequence.
55 1866 pSAC35:teprotide.HSA Teprotide fused upstream of mature HSA. pSAC35 271 55 487 751 752
56 1867 pSAC35:HSA.teprotide. Teprotide fused downstream of FL HSA. pSAC35 272 56 488 753 754 HSA
57 1889 pC4:HSA.PTH.S1-F34 PTH(1-34) fused downstream of HSA. pC4 273 57 489 755 756 HSA
58 1891 pEE12:HSA.sTR6 Soluble mature TR6 fused downstream of HSA. pEE12.1 274 58 490 757 758 HSA
59 1892 pEE12:sTR6.HSA Synthetic full length TR6 fused upstream of mature pEE12.1 275 59 491 759 760 TR6
HSA.
60 1906 pC4:PTH.S1-F34.HSA Amino acids S1 to F34 of PTH fused upstream of pC4 276 60 492 761 762 MPIF
(junctioned) mature HSA and downstream of MPIF leader
sequence. There are two cloning junction amino
acids (T, S) between PTH and HSA.
61 1907 pC4:HSA.PTH.S1-F34 Amino acids S1 to F34 fused downstream of FL pC4 277 61 493 763 764 HSA
(junctioned) HSA. The last C-terminal amino acid (L) residue
is missing for HSA in the cloning junction between
HSA and PTH.
62 1912 pC4:sTR6.HSA Synthetic full length TR6 fused upstream of mature pC4 278 62 494 765 766 Native TR6
HSA. leader
63 1913 pC4:HSA.synTR6.V30-H300 Amino acids V30 to H300 of synthetic TR6 pC4 279 63 495 767 768 HSA
(seamless) (shown as V1 to H271 of SEQ ID NO: 495) fused
downstream of full-length HSA.
64 1914 pC4:PTH.S1-F34.HSA Amino acids S1 to F34 of PTH fused downstream pC4 280 64 496 769 770 MPIF
(seamless) of MPIF leader sequence and upstream of mature
HSA.
65 1916 pC4:HSA.KGF2D28.A63-S208 Amino acids A63 to S208 of full length KGF2 pC4 281 65 497 771 772 HSA
fused downstream of HSA.
66 1917 pC4:KGF2D28.A63-S208: Amino acids A63 to S208 of KGF2 fused upstream pC4 282 66 498 773 774 HSA/kex2
HSA of mature HSA.
67 1925 pcDNA3.EPO M1-D192. Amino acids M1 to D192 of EPO variant (where pcDNA3 283 67 499 775 776 Native EPO
HSA glycine at amino acid 140 has been replaced with leader peptide
an arginine) fused upstream of HSA. D192 of EPO
and D1 of mature HSA are the same amino acids in
this construct.
68 1926 pcDNA3:SPHSA.EPO A28-D192 Amino acids A28 to D192 of EPO variant (where pcDNA3 284 68 500 777 778 MPIF
glycine at amino acid 140 has been replaced with
an arginine) fused upstream of mature HSA and
downstream of the MPIF leader peptide.
69 1932 pEE12.1:HSA.PTH.S1-F34 Amino acids 1 to 34 of PTH fused downstream of pEE12.1 285 69 501 779 780 HSA
full length HSA.
70 1933 pSAC35:HCC-1.T20-N93: Amino acids T20 to N93 of HCC-1 fused upstream pSAC35 286 70 502 781 782 HSA/kex2
HSA of mature HSA and downstream of the HSA/kex2
leader sequence.
71 1934 pSAC35:HCC-1C.O.T20-N93: Amino acids T20 to N93 of HCC-1 fused upstream pSAC35 287 71 503 783 784 HSA/kex2
HSA of mature HSA and downstream of the HSA/kex2
leader sequence. DNA sequence is codon
optimized for yeast expression.
72 1938 pEE12.1:PTH.S1-F34.HSA Amino acids S1 to F34 of PTH fused upstream of pEE12.1 288 72 504 785 786 MPIF
mature HSA and downstream of MPIF leader
sequence.
73 1941 pC4:HSA/PTH84 PTH fused downstream of full length HSA. The pC4 289 73 505 787 788 HSA
(junctioned) last amino acid of HSA (Leu) has been deleted.
74 1947 pSAC35:d8HCC-1.G28-N93: Amino acids G28 to N93 of HCC-1 fused upstream pSAC35 290 74 506 789 790 HSA/kex2
HSA of mature HSA and downstream of HSA/kex2
leader sequence.
75 1948 pSAC35:d8HCC-1C.O.G28-N93: Amino acids G28 to N93 of HCC-1 fused upstream pSAC35 291 75 507 791 792 HSA/kex2
HSA of mature HSA and downstream of HSA/kex2
leader sequence. DNA sequence is codon
optimized for yeast expression.
76 1949 pC4:PTH.S1-Q84/HSA PTH fused downstream of the MPIF leader pC4 292 76 508 793 794 MPIF
(junctioned) sequence and upstream of mature HSA. There are
two additional amino acids between PTH84 and
HSA as a result of the cloning site.
77 1952 pcDNA3.1:IL2.HSA Full length human IL-2, having a Cysteine to pCDNA3.1 293 77 509 795 796 Native IL-2
Serine mutation at amino acid 145, fused upstream leader
of mature HSA.
78 1954 pC4:IL2.HSA Full length human IL-2, having a Cysteine to pC4 294 78 510 797 798 Native IL-2
Serine mutation at amino acid 145, fused upstream leader
of mature HSA.
79 1955 pSAC35:t9HCC-1.G28-N93: Amino acids G28 to N93 of HCC-1 fused upstream pSAC35 295 79 511 799 800 HSA/kex2
spcHSA of a 16 amino acid spacer and mature HSA and
downstream of HSA/kex2 leader sequence.
80 1956 pSAC35:HSA.scFv116A01 Single chain BLyS antibody fused downstream of pSAC35 296 80 512 801 802 HSA/kex2
HSA with HSA/kex2 leader sequence. This
construct also contains a His tag at the 3′ end.
81 1966 pC4:EPO.M1-D192.HSA Amino acids M1 to D192 of EPO variant (where pC4 297 81 513 Native EPO
Construct is also named glycine at amino acid 140 has been replaced with leader peptide
pC4:EPOM1-D192.HSA an arginine) fused upstream of mature HSA.
82 1969 pC4:MPIFsp.HSA.EPO.A28-D192 Amino acids A28 to D192 of EPO variant (where pC4 298 82 514 MPIF
glycine at amino acid 140 has been replaced with
an arginine) fused downstream of MPIF leader
sequence and upstream of mature HSA.
83 1980 pC4:EPO.A28-D192.HSA Amino acids A28 to D192 of EPO variant (where pC4 299 83 515 803 804 HSA
glycine at amino acid 140 has been replaced with
an arginine) fused downstream of the HSA leader
peptide and upstream of mature HSA.
84 1981 pC4.HSA-EPO.A28-D192. Amino acids A28 to D192 of EPO variant (where pC4 300 84 516 805 806 HSA
glycine at amino acid 140 has been replaced with
an arginine) fused downstream of the full length
HSA.
85 1989 pSAC35:activeAC2inhibitor: Active inhibitor of ACE2 (DX512) fused upstream pSAC35 301 85 517 807 808 HSA/kex2
HSA of mature HSA and downstream of HSA/kex2
leader sequence.
86 1994 pEE12.1.HSA-EPO.A28-D192. Amino acids A28 to D192 of EPO variant (where pEE12.1 302 86 518 HSA
glycine at amino acid 140 has been replaced with
an arginine) fused downstream of full length HSA.
87 1995 pEE12.1:EPO.A28-D192. Amino acids A28 to D192 of EPO variant (where pEE12.1 303 87 519 HSA
HSA glycine at amino acid 140 has been replaced with
an arginine) fused downstream of the HSA leader
peptide and upstream of mature HSA.
88 1996 pEE12.1:MPIFsp.HSA.EPO. Amino acids A28 to D192 of EPO variant (where pEE12.1 304 88 520 MPIF
A28-D192 glycine at amino acid 140 has been replaced with
an arginine) fused downstream of MPIF leader
sequence and upstream of mature HSA.
89 1997 pEE12.1:EPO M1-D192.HSA Amino acids M1 to D192 of EPO variant (where pEE12.1 305 89 521 Native EPO
glycine at amino acid 140 has been replaced with leader
an arginine) fused upstream of mature HSA.
90 1998 pC4:CKB1.G28-N93.HSA Amino acids G28 to N93 of CkBeta1 fused pC4 306 90 522 809 810 HSA
upstream of mature HSA and downstream of the
HSA leader sequence.
91 2000 pSAC35:HSA:activeAC2inhibitor Active inhibitor of ACE2 (DX512) fused pSAC35 307 91 523 811 812 HSA
downstream of HSA.
92 2001 pSAC35:inactiveAC2inhibitor: Inactive inhibitor of ACE2 (DX510) fused pSAC35 308 92 524 813 814 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
93 2002 pSAC35:HSA.inactiveAC2inhibitor Inactive inhibitor of ACE2 (DX510) fused pSAC35 309 93 525 815 816 HSA
downstream of HSA.
94 2011 pC4:IFNb-HSA Full length IFNb fused upstream of mature HSA. pC4 310 94 526 817 818 Native IFNb
leader
95 2013 pC4:HSA-IFNb.M22-N187 Amino acids M22 to N187 of IFNb (fragment pC4 311 95 527 HSA
shown as amino acids M1 to N166 of SEQ ID
NO: 527) fused downstream of HSA.
96 2016 pC4:TR1.M1-L401.HSA Amino acids M1 to L401 of TR1 fused upstream of pC4 312 96 528 819 820 Native TR1
mature HSA. Native TR1 signal sequence used. A
Kozak sequence was added.
97 2017 pC4:HSA.TR1.E22-L401 Amino acids E22 to L401 of TR1 fused pC4 313 97 529 821 822 HSA
downstream of HSA.
98 2021 pC4:PTH.S1-Q84/HSA Amino acids 1-84 of PTH fused upstream of pC4 314 98 530 823 824 HSA
(seamless) mature HSA and downstream of native HSA leader
sequence.
99 2022 pEE12.1:PTH.S1-Q84.HSA Amino acids 1-84 of PTH fused upstream of pEE12.1 315 99 531 HSA
mature HSA and downstream of native HSA leader
sequence.
100 2023 pSAC35.PTH.S1-Q84.HSA Amino acids 1-84 of PTH fused upstream of pSAC35 316 100 532 825 826 HSA/kex2
mature HSA and downstream of HSA/kex2 leader
sequence.
101 2025 pSAC35:teprotide.spacer.HSA Teprotide fused upstream of a linker and mature pSAC35 317 101 533 827 828
HSA.
102 2026 pSAC35:HSA.spacer.teprotide Teprotide fused downstream of HSA and a linker. pSAC35 318 102 534 829 830 HSA
103 2030 pSAC35.ycoIL-2.A21-T153. Amino acids A21 to T153 of IL-2 fused upstream pSAC35 319 103 535 831 832 HSA/kex2
HSA of mature HSA and downstream of HSA/kex2
leader sequence. DNA encoding IL-2 has been
codon optimized.
104 2031 pSAC35.HSA.ycoIL-2.A21-T153 Amino acids A21 to T153 of IL-2 fused pSAC35 320 104 536 833 834 HSA/kex2
downstream of HSA with the HSA/kex2 leader
sequence. DNA encoding IL-2 has been codon
optimized.
105 2047 pC4HSA:SP.EPO A28-D192. Amino acids A28 to D192 of EPO variant (where pSAC35 321 105 537 835 836 MPIF
HSA glycine at amino acid 140 has been replaced with
an arginine) fused upstream of mature HSA and
downstream of MPIF leader peptide.
106 2053 pEE12:IFNb-HSA Full length IFNb fused upstream of mature HSA. pEE12.1 322 106 538 Native IFNb
also named pEE12.1:IFNβ- leader
HSA
107 2054 pEE12:HSA-IFNb Mature IFNb fused downstream of HSA. pEE12.1 323 107 539 HSA
108 2066 pC4:GM-CSF.M1-E144.HSA Amino acids M1 to E144 of GM-CSF fused pC4 324 108 540 837 838 Native GM-
upstream of mature HSA. CSF
109 2067 pC4:HSA.GM-CSF.A18-E144 Amino acids A18 to E144 of GM-CSF fused pC4 325 109 541 839 840 HSA
downstream of HSA.
110 2085 pEE12.1:TR1.M1-L401.HSA Amino acids M1 to L401 of TR1 fused upstream of pEE12.1 326 110 542 Native TR-1
mature HSA.
111 2086 pEE12.1:HSA.TR1.E22-L401 Amino acids E22 to L401 (fragment shown as pEE12.1 327 111 543 HSA
amino acids E1 to L380 of SEQ ID NO: 543) of
TR1 fused downstream of HSA.
112 2095 pC4:HSA-BLyS.A134 Amino acids A134 to L285 of BLyS fused pC4 328 112 544 841 842 HSA
downstream of HSA.
113 2096 pC4:sp.BLyS.A134-L285. Amino acids A134 to L285 of BLyS (fragment pC4 329 113 545 843 844 Native CKβ8
HSA shown as amino acids A1 to L152 of SEQ ID
NO: 545) fused upstream of mature HSA and
downstream of the CKb8 signal peptide.
114 2101 pcDNA3:SP.Ck7 Q22-A89. N-terminal Methionine fused to amino acids Q22 pcDNA3 330 114 546 845 846 MPIF
HSA. to A89 of Ckβ7 fused upstream of mature HSA and
downstream of MPIF signal peptide.
115 2102 pEE12.1:SP.EPO A28-D192. Amino acids A28 to D192 of EPO variant (where pEE12.1 331 115 547 MPIF
HSA glycine at amino acid 140 has been replaced with
an arginine) fused upstream of mature HSA and
downstream of MPIF leader peptide.
116 2129 pC4:TR2.M1-A192.HSA Amino acids M1-A192 of TR2 fused upstream of pC4 332 116 548 847 848 Native TR2
HSA.
117 2137 pSAC35.MDC.G25-Q93. Amino acids G25 to Q93 of MDC fused upstream pSAC35 333 117 549 849 850 HSA/kex2
HSA. of mature HSA and downstream of HSA/kex2
leader sequence.
118 2141 HSA-CK-Beta4 Full length CK-beta4 fused downstream of HSA. pSAC35 334 118 550 851 852 HSA
119 2146 pC4:Leptin.HSA Full length Leptin fused upstream of mature HSA. pC4 335 119 551 853 854 Native leptin
120 2181 pC4:HSA.IL1Ra(R8-E159) Amino acids R8 to E159 of IL1Ra (plus an added pC4 336 120 552 855 856 HSA
methionine at N-terminus) fused downstream of
HSA.
121 2182 pC4:MPIFsp(M1-A21). Amino acids R8 to E159 of IL1Ra (plus an added pC4 337 121 553 857 858 MPIF
IL1Ra(R8-E159).HSA methionine at N-terminus) fused downstream of the
MPIF leader sequence and upstream of mature
HSA.
122 2183 pSAC35:HSA.IL1Ra(R8-E159) Amino acids R8 to E159 of IL1Ra (plus an added pSAC35 338 122 554 859 860 HSA
methionine at N-terminus) fused downstream of
HSA.
123 2184 pC4:HSA.Leptin.V22-C166 Amino acids V22 to C167 of Leptin fused pC4 339 123 555 861 862 HSA
downstream of HSA.
124 2185 pSAC35:IL1Ra(R8-E159). Amino acids R8 to E159 of IL1Ra (plus an added pSAC35 340 124 556 863 864 HSA/kex2
HSA methionine at N-terminus) fused upstream of
mature HSA and downstream of HSA/kex2 leader
sequence.
125 2186 pSAC35:Leptin.V22-C166. Amino acids V22 to C167 of Leptin fused pSAC35 341 125 557 865 866 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
126 2187 pSAC35:HSA.Leptin.V22-C166 Amino acids V22 to C167 of Leptin fused pSAC35 342 126 558 867 868 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence.
127 2226 pcDNA3(+):TREM-1(21-202)- Amino acids A21 to P202 of TREM-1 fused pCDNA3.1 343 127 559 869 870 MPIF
HSA upstream of mature HSA and downstream of the
MPIF leader sequence.
128 2230 pC4:TREM-1.M1-P202.HSA Amino acids M1 to P202 of TREM-1 fused pC4 344 128 560 871 872 Native
upstream of mature HSA. TREM-1
129 2240 pC4:SP.Ck7 Q22-A89.HSA. N-terminal Methionine fused to amino acids Q22 pC4 345 129 561 873 874 MPIF
to A89 of Ckβ7 fused upstream of mature HSA and
downstream of the MPIF leader sequence.
Contains a linker sequence between Ckβ7 and
HSA.
130 2241 pC4:HSA.Ck7metQ22-A89. N-terminal Methionine fused to amino acids Q22 pC4 346 130 562 875 876 HSA/kex2
to A89 of Chemokine beta 7 (Ckbeta 7 or CK7)
fused downstream of HSA with HSA/kex2 leader
sequence. Contains a linker sequence between
CkB7 and HSA.
131 2244 pC4.HCNCA73.HSA HCNCA73 fused upstream of mature HSA. pC4 347 131 563 877 878 HCNCA73
132 2245 pScNHSA:CK7.Q22-A89 Amino acids Q22 to A89 of Ckβ7 fused pScNHSA 348 132 564 879 880 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence. Contains a linker sequence between
Ckβ7 and HSA.
133 2246 pScCHSA.CK7metQ22-A89 N-terminal Methionine fused to amino acids Q22 pScCHSA 349 133 565 881 882 HSA/kex2
to A89 of Ckβ7 fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
134 2247 pSAC35:CK7metQ22-A89. N-terminal Methionine fused to amino acids Q22 pSAC35 350 134 566 883 884 HSA/kex2
HSA. to A89 of Ckβ7 fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
135 2248 pSAC35:HSA.CK7metQ22-A89. N-terminal Methionine fused to amino acids Q22 pSAC35 351 135 567 885 886 HSA/kex2
to A89 of Ckβ7 fused downstream of HSA with
HSA/kex2 leader sequence. Contains a linker
sequence between Ckβ7 and HSA.
136 2249 pSAC35:IFNa2-HSA Mature IFNa2 fused upstream of mature HSA and pSAC35 352 136 568 887 888 HSA/kex2
also named: pSAC23:IFNα2- downstream of HSA/kex2 leader sequence.
HSA
137 2250 pSAC35:HSA.INSULIN(GYG) Mature Insulin wherein the C-peptide is replaced pSAC35 353 137 569 889 890 HSA
also named: by the C-domain of IGF-1 fused downstream of
pSAC35.HSA.INSULING(GYG). HSA. DNA encoding Insulin was codon optimized.
F1-N62
138 2251 pScCHSA:VEGF2.T103-R227. Amino acids T103 to R227 of VEGF2 fused pScCHSA 354 138 570 891 892 HSA/kex2
upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
139 2252 pScNHSA:VEGF2.T103-R227. Amino acids T103 to R227 of VEGF2 fused pScNHSA 355 139 571 893 894 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence.
140 2255 pSAC35:INSULIN(GYG).HSA Mature Insulin wherein the C-peptide is replaced pSAC35 356 140 572 895 896 HSA/kex2
also named by the C-domain of IGF-1 fused upstream of
pSAC35.INSULING(GYG).F1-N62. mature HSA and downstream of HSA/kex2 leader.
HSA DNA encoding Insulin was codon optimized.
141 2256 pSAC35:VEGF2.T103-R227. Amino acids T103 to R227 of VEGF2 fused pSAC35 357 141 573 897 898 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
142 2257 pSAC35:HSA.VEGF2.T103-R227 Amino acids T103 to R227 of VEGF-2 fused pSAC35 358 142 574 899 900 HSA/kex2
downstream of HSA with HSA/kex2 leader
sequence.
143 2271 pEE12.1:HCHNF25M1-R104. Amino acids M1 to R104 of HCHNF25 fused pEE12.1 359 143 575 Native
HSA upstream of mature HSA. HCHNF25
144 2276 pSAC35:HSA.INSULIN(GGG) Mature Insulin wherein the C-peptide is replaced pSAC35 360 144 576 901 902 HSA
also named: by a synthetic linker fused downstream of HSA.
pSAC35.HSA.INSULING(GGG). DNA encoding Insulin was codon optimized.
F1-N58
145 2278 pSAC35:insulin(GGG).HSA Mature Insulin wherein the C-peptide is replaced pSAC35 361 145 577 903 904 HSA/kex2
by a synthetic linker fused downstream of
HSA/kex2 leader and upstream of mature HSA.
DNA encoding Insulin was codon optimized.
146 2280 pC4:HCHNF25.HSA HCHNF25 fused upstream of mature HSA. pC4 362 146 578 905 906 Native
HCHNF25
147 2283 pScCHSA:EPOcoA28-D192. Amino acids A28 to D192 of EPO variant (where pScCHSA 363 147 579 907 908 HSA/kex2
51N/Q, 65N/Q, glycine at amino acid 140 has been replaced with
110N/Q EPO an arginine) are fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
Glycosylation sites at amino acids 51, 65 and 110
are mutated from N to Q residue. DNA encoding
EPO is codon optimized.
148 2284 pScNHSA:EPOcoA28-D192. Amino acids A28 to D192 of EPO variant (where pScNHSA 364 148 580 909 910 HSA/kex2
51N/Q, 65N/Q, glycine at amino acid 140 has been replaced with
110N/Q EPO an arginine) fused downstream of mature HSA and
HSA/kex2 leader sequence. Glycosylation sites at
amino acids 51, 65 and 110 are mutated from N to
Q residue. DNA encoding EPO is codon
optimized.
149 2287 pSAC35:EPOcoA28-D192. Amino acids A28 to D192 of EPO variant (where pSAC35 365 149 581 911 912 HSA/kex2
51N/Q, 65N/Q, 110N/Q. glycine at amino acid 140 has been replaced with
HSA. an arginine) fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
Glycosylation sites at amino acid 51, 65 and 110
are mutated from N to Q residue. DNA encoding
EPO is codon optimized.
150 2289 pSAC35:HSA.EPOcoA28-D192. Amino acids A28 to D192 of EPO variant (where pSAC35 366 150 582 913 914 HSA/kex2
51N/Q, 65N/Q, 110N/Q. glycine at amino acid 140 has been replaced with
an arginine) fused downstream of mature HSA and
HSA/kex2 leader sequence. Glycosylation sites at
amino acid 51, 65 and 110 are mutated from N to
Q residue. DNA encoding EPO is codon
optimized.
151 2294 pC4:EPO.R140G.HSA Amino acids M1-D192 of EPO fused upstream of pC4 367 151 587 915 916 Native EPO
also named mature HSA. The EPO sequence included in
pC4.EPO.R1406.HSA construct 1997 was used to generate this construct,
mutating arginine at EPO amino acid 140 to
glycine. This mutated sequence matches the
wildtype EPO sequence.
152 2295 pSAC35:humanresistin.K19-P108: Amino acids K19 to P108 of Resistin fused pSAC35 368 152 584 917 918 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
153 2296 pSAC35:HSA:humanresistin. Amino acids K19 to P108 of Resistin fused pSAC35 369 153 585 919 920 HSA
K19-P108 downstream of HSA.
154 2297 pSAC35:humanresistin.K19-P108. Amino acids K19 to P108 of Resistin fused pSAC35 370 154 586 921 922 HSA/kex2
stop:HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence. Includes two stops at 3′
end for termination of translation before the HSA.
155 2298 pEE12.1:EPO.R140G.HSA Amino acids M1 to D192 of EPO fused upstream pEE12.1 371 155 587 923 924 Native EPO
of mature HSA. The EPO sequence included in
construct 1997 was used to generate this construct,
mutating arginine at EPO amino acid 140 to
glycine. This mutated sequence matches the
wildtype EPO sequence.
156 2300 pC4:humanresistin.M1-P108: Amino acids M1 to P108 of Resistin fused pC4 372 156 588 925 926 Native
HSA upstream of mature HSA. resistin
157 2309 pEE12.1:humanresistin.M1-P108: Amino acids M1 to P108 of Resistin fused pEE12.1 373 157 589 927 Native
HSA upstream of mature HSA. resistin
158 2310 pc4:EPOco.M1-D192.HSA Amino acids M1 to D192 of EPO variant fused pC4 374 158 590 928 929 Native EPO
upstream of mature HSA. DNA encoding EPO is
codon optimized. The EPO sequence included in
construct 1997 was used to generate this construct,
mutating arginine at EPO amino acid 140 to
glycine. This mutated sequence matches the
wildtype EPO sequence.
159 2311 pC4:EPO.M1-G27. Amino acids M1 to D192 of EPO fused upstream pC4 375 159 591 930 931 Native EPO
EPOco.A28-D192.HSA of mature HSA. DNA encoding only EPO portion
is codon optimized. The EPO sequence included
in construct 1997 was used to generate this
construct, mutating arginine at EPO amino acid
140 to glycine. This mutated sequence matches the
wildtype EPO sequence.
160 2320 pC4:HCHNF25M1-R104. Amino acids M1 to R104 of HCHNF25 fused pC4 376 160 592 932 933 Native
HSA upstream of mature HSA. HCHNF25
161 2325 pC4.EPO:M1-D192. Amino acids M1 to D192 of EPO fused upstream pC4 377 161 593 Native EPO
HSA.Codon opt. of mature HSA. DNA encoding EPO is codon
optimized.
162 2326 pEE12.1.EPO:M1-D192. Amino acids M1 to D192 of EPO fused upstream pEE12.1 378 162 594 Native EPO
HSA.Codon opt. of mature HSA. DNA encoding EPO is codon
optimized.
163 2328 pC4:HLDOU18.K23-R429. Amino acids K23 to R429 of HLDOU18 fused pC4 379 163 595 934 935 HSA
HSA upstream of mature HSA and downstream of native
HSA leader sequence.
164 2330 CK-Beta4-HSA Full length Ckbeta4 fused upstream of mature pSAC35 380 164 596 936 937 Native CKβ4
HSA.
165 2335 pC4:MPIFsp.ck{b}4D31-M96. Amino acids D31 to M96 of Ckbeta4 fused pC4 381 165 597 938 939 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
166 2336 pC4:MPIFsp.ck{b}4G35-M96. Amino acids G35 to M96 of Ckbeta4 fused pC4 382 166 598 940 941 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
167 2337 pC4:MPIFsp.ck{b}4G48-M96. Amino acids G48 to M96 of Ckbeta4 fused pC4 383 167 599 942 943 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
168 2338 pC4:MPIFsp.ck{b}4A62-M96. Amino acids A62 to M96 of Ckbeta4 fused pC4 384 168 600 944 945 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
169 2340 pC4:HSA.HLDOU18.K23-R429 Amino acids K23 to R429 of HLDOU18 fused pC4 385 169 601 946 947 HSA
downstream of HSA.
170 2343 pSAC35.INV-IFNA2.HSA Mature Interferon alpha2 fused upstream of mature pSAC35 386 170 602 948 949 invertase
HSA and downstream of invertase signal peptide.
171 2344 pC4.SpIg.EPO:A28-D192. Amino acids A28 to D192 of EPO fused upstream pC4 387 171 603 950 951 Mouse Ig
HSA.Codon opt. of mature HSA and downstream of mouse Ig leader leader
sequence. DNA encoding EPO is codon optimized.
172 2348 pC4:MPIFsp.ck{b}4G57-M96. Amino acids G57 to M96 of Ckbeta4 fused pC4 388 172 604 952 953 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
173 2350 pC4:MPIFsp.HLDOU18(S320-R429). Amino acids S320 to R429 of HLDOU18 fused pC4 389 173 605 954 955 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
174 2351 pC4:HSA.HLDOU18(S320-R429) Amino acids S320 to R429 of HLDOU18 fused pC4 390 174 606 956 957 HSA
downstream of HSA.
175 2355 pSAC35:MATalpha.d8ckbeta1. Amino acids G28 to N93 of Ckbeta1 fused pSAC35 391 175 607 958 959 MFα-1
G28-N93:HSA upstream of mature HSA and downstream of the
yeast mating factor alpha leader sequence.
176 2359 pEE12:HLDOU18.K23-R429. Amino acids K23 to R429 of HLDOU18 fused pEE12.1 392 176 608 HSA
HSA upstream of mature HSA and downstream of native
HSA leader sequence.
177 2361 pC4:HRDFD27:HSA HRDFD27 fused upstream of mature HSA. pC4 393 177 609 960 961 Native
HRDFD27
178 2362 pEE12:HSA.HLDOU18.K23-R429 Amino acids K23 to R429 of HLDOU18 fused pEE12.1 394 178 610 HSA
downstream of HSA.
179 2363 pC4GCSF.HSA.EPO.A28-D192 Amino acids M1 to P204 of GCSF fused upstream pC4 395 179 611 Native GCSF
of mature HSA which is fused upstream of amino
acids A28 to D192 of EPO variant (where amino
acid 140 of EPO is mutated from glycine to
arginine.)
180 2365 pEE12.1.HCNCA73HSA HCNCA73 is fused upstream of mature HSA. pEE12.1 396 180 612 962 963 Native
HCNCA73
181 2366 pSAC35.MAF-IFNa2.HSA Mature IFNa2 fused upstream of mature HSA and PSAC35 397 181 613 964 965 MFα-1
downstream of yeast mating factor alpha leader
sequence.
182 2367 pEE12.MPIFsp.HLDOU18.S320-R429. Amino acids S320 to R429 of HLDOU18 fused pEE12.1 398 182 614 966 967 MPIF
HSA upstream of mature HSA and downstream of MPIF
leader sequence.
183 2369 pC4:HLDOU18.HSA Amino acids M1 to R429 of HLDOU18 fused pC4 399 183 615 968 969 Native
upstream of mature HSA. HLDOU18
184 2370 pEE12:HLDOU18.HSA Amino acids M1 to R429 of HLDOU18 fused pEE12.1 400 184 616 Native
upstream of mature HSA. HLDOU18
185 2373 pC4.GCSF.HSA.EPO.A28-D192. Amino acids M1 to P204 of GCSF is fused pC4 401 185 617 Native GCSF
R140G upstream of mature HSA which is fused upstream
of amino acids A28 to D192 of EPO, wherein
amino acid 140 is glycine. The EPO sequence
included in construct 1997 was used to generate
this construct, mutating arginine at EPO amino
acid 140 to glycine. This mutated sequence
matches the wildtype EPO sequence.
186 2381 pC4:HSA-IFNa2(C17-E181) Amino acids C17 to E181 of IFNa2 (fragment pC4 402 186 618 970 971 HSA
shown as amino acids C1 to E165 of SEQ ID
NO: 618) fused downstream of HSA.
187 2382 pC4:IFNa2-HSA IFNa2 fused upstream of mature HSA. pC4 403 187 619 972 973 Native IFNα2
leader
188 2387 pC4:EPO(G140)-HSA- Amino acids M1-D192 of EPO fused upstream of pC4 404 188 620 Native EPO
GCSF.T31-P204 mature HSA which is fused upstream of amino
acids T31 to P204 of GCSF.
189 2407 pC4:HWHGZ51.M1-N323. Amino acids M1 to N323 of HWHGZ51 fused pC4 405 189 621 974 975 Native
HSA upstream of mature HSA. HWHGZ51
190 2408 pEE12.1:HWHGZ51.M1-N323. Amino acids M1 to N323 of HWHGZ51 fused pEE12.1 406 190 622 976 977 Native
HSA upstream of mature HSA. HWHGZ51
191 2410 pSAC35INV:IFNa-HSA Mature IFNa2 fused downstream of the invertase pSAC35 407 191 623 978 979 invertase
signal peptide and upstream of mature HSA.
192 2412 pSAC35:delKEX.d8ckbeta1. Amino acids G28 to N93 of Ckbeta1 fused pSAC35 408 192 624 980 981 HSA minus
G28-N93:HSA downstream of the HSA signal sequence (with the the KEX site
KEX site deleted - last 6 amino acids of the leader)
and upstream of mature HSA.
193 2414 pC4.EPO:M1-D192copt. Amino acids M1 to D192 of EPO fused upstream pC4 409 193 625 982 983 Native EPO
HSA.GCSF.T31-P204 of mature HSA which is fused upstream of amino
also named: acids T31 to P204 of GCSF. DNA encoding EPO
pC4.EPO:M1-D192copt. has been codon optimized.
HAS.GCSF.T31-P204
194 2428 pN4:PTH.S1-Q84/HSA Amino acids S1 to Q84 of PTH fused upstream of pN4 410 194 626 HSA
mature HSA and downstream of the native HSA
leader sequence.
195 2441 pEE12.EPO:M1-D192copt. Amino acids M1 to D192 of EPO fused upstream pEE12.1 409 196 628 EPO leader
HSA.GCSF.T31-P204 of mature HSA which is fused upstream of amino
also named: acids T31 to P204 of GCSF. DNA encoding EPO
pEE12.EPO:M1-D192copt. has been codon optimized.
HAS.GCSF.T31-P204
196 2447 pC4:HSA.humancalcitonin.C1-G33 Amino acids C98 to G130 of SEQ ID NO: 629 pC4 413 197 629 986 987 HSA
fused downstream of HSA.
197 2448 pSAC35:GLP-1(7-36).HSA Amino acids H98 to R127 of preproglucagon (SEQ pSAC35 414 198 630 988 989 HSA/kex2
ID NO: 630) (hereinafter this specific domain will
be referred to as “GLP-1(7-36)”) is fused upstream
of mature HSA and downstream of HSA/kex2
leader sequence.
198 2449 pSAC35:INV.d8CKB1.G28-N93: Amino acids G28 to N93 of Ckbeta1 fused pSAC35 415 199 631 990 991 Invertase
HSA downstream of the invertase signal peptide and
upstream of mature HSA.
199 2455 pSAC35:HSA.GLP-1(7-36) GLP-1(7-36) is fused downstream of mature HSA pSAC35 416 200 632 992 993 HSA/kex2
and HSA/kex2 leader sequence.
200 2456 pSAC35:GLP-1(7- Amino acids H98 to R127 of Preproglucagon (SEQ pSAC35 417 201 633 994 995 HSA/kex2
36(A8G)).HSA ID NO: 633)(also referred to as “GLP-1(7-36)”) is
mutated at amino acid 99 of SEQ ID NO: 633 to
replace the alanine with a glycine. This particular
GLP-1 mutant will be hereinafter referred to as
“GLP-1(7-36(A8G))” and corresponds to the
sequence shown in SEQ ID NO: 1808. GLP-1(7-
36(A8G)) is fused upstream of mature HSA and
downstream of HSA/kex2 leader sequence.
201 2457 pSAC35:HSA.GLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is fused pSAC35 418 202 634 996 997 HSA/kex2
36(A8G)) downstream of mature HSA and HSA/kex2 leader
sequence.
202 2469 pSAC35:HSA.exendin.H48-S86 Amino acids H48 to S86 of Exendin fused pSAC35 419 203 635 HSA
downstream of full length HSA.
203 2470 pSAC35:Exendin.H48-S86. Amino acids H48 to S86 of Exendin fused pSAC35 420 204 636 HSA/kex2
HSA upstream of mature HSA and downstream of
HSA/kex2 leader sequence.
204 2473 pC4.HLDOU18:HSA:S320-R429 M1-R319 of HLDOU18 (containing the furin site pC4 421 205 637 998 999 Native
RRKR) followed by residues ‘LE’ followed by HLDOU18
mature HSA followed by ‘LE’ and amino acids
S320 through R429 of HLDOU18 (fragment
shown as SEQ ID NO: 637).
205 2474 pSAC35.MDC.P26-Q93.HSA Amino acids P26 to Q93 of MDC fused pSAC35 422 206 638 1000 1001 HSA/kex2
downstream of the HSA/kex2 leader and upstream
of mature HSA.
206 2475 pSAC35.MDC.M26-Q93. Amino acids Y27 to Q93 of MDC with an N- pSAC35 423 207 639 1002 1003 HSA/kex2
HSA terminal methionine, fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
207 2476 pSAC35.MDC.Y27-Q93. Amino acids Y27 to Q93 of MDC fused pSAC35 424 208 640 1004 1005 HSA/kex2
HSA downstream of the HSA/kex2 leader and upstream
of mature HSA.
208 2477 pSAC35.MDC.M27-Q93. Amino acids G28 to Q93 of MDC with an N- pSAC35 425 209 641 1006 1007 HSA/kex2
HSA terminal methionine, fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
209 2489 pSAC35:HSA.C17.A20-R136 Amino acids A20 to R136 of C17 fused pSAC35 426 210 642 1008 1009 HSA/kex2
downstream of mature HSA with HSA/kex2 leader
sequence.
210 2490 pSAC35:C17.A20-R136.HSA Amino acids A20 to R136 of C17 fused pSAC35 427 211 643 1010 1011 HSA/kex2
downstream of the HSA/kex2 leader and upstream
of mature HSA.
211 2492 pC4.IFNb(deltaM22).HSA Mutant full length INFbeta fused upstream of pC4 428 212 644 Native IFNβ
mature HSA. First residue of native, mature leader
IFNbeta (M22) has been deleted.
212 2498 pC4:HSA.KGF2D60.G96-S208 Amino acids G96 to S208 of KGF-2 fused pC4 429 213 645 1012 1013 HSA
downstream of HSA.
213 2499 pC4:KGF2D60.G96-S208: Amino acids G96 to S208 of KGF2 fused upstream pC4 430 214 646 1014 1015 HSA
HSA of mature HSA and downstream of the HSA signal
peptide.
214 2501 pSAC35:scFvI006D08.HSA BLyS antibody fused upstream of mature HSA and pSAC35 431 215 647 1016 1017 HSA/kex2
downstream of HSA/kex2 signal peptide.
215 2502 pSAC35:scFvI050B11.HSA BLyS antibody fused upstream of mature HSA and pSAC35 432 216 648 1018 1019 HSA/kex2
downstream of HSA/kex2 leader sequence.
216 2513 pC4:HSA.salmoncalcitonin.C1-G33 C1 through G33 of salmon calcitonin fused pC4 1513 1345 1681 1854 1855 HSA
downstream of full length HSA.
217 2515 pC4:HDPBQ71.M1-N565. M1 through N565 of HDPBQ71 fused upstream of pC4 1514 1346 1682 1856 1857 Native
HSA mature HSA HDPBQ71
218 2529 pC4:TR1.M1-K194.HSA Amino acids M1 to K194 of TR1 (including native pC4 1223 1208 1238 1253 1254 Native TR1
signal sequence) fused upstream of mature HSA.
219 2530 pC4:TR1.M1-Q193.HSA Amino acids M1 to Q193 of TR1 (including native pC4 1224 1209 1239 1255 1256 Native TR1
signal sequence) fused upstream of mature HSA.
220 2531 pC4:TR1.M1-E203.HSA Amino acids M1 to E203 of TR1 (including native pC4 1225 1210 1240 1257 1258 Native TR1
signal sequence) fused upstream of mature HSA.
221 2532 pC4:TR1.M1-Q339.HSA Amino acids M1 to Q339 of TR1 (including native pC4 1226 1211 1241 1259 1260 Native TR1
signal sequence) fused upstream of mature HSA.
222 2545 pEE12.1:HDPBQ71.M1-N565. M1 through N565 of HDPBQ71 fused upstream of pEE12.1 1515 1347 1683 Native
HSA mature HSA HDPBQ71
223 2552 pSAC35:KGF2delta33.S69-S208. Amino acids S69 through S208 of KGF2 fused pScCHSA 1516 1348 1684 1858 1859 HSA/kex2
HSA upstream of HSA.
224 2553 pSAC35:HSA.KGF2delta33. HSA/kex2 signal peptide followed by HSA peptide pScNHSA 1517 1349 1685 1860 1861 HSA/kex2
S69-S208 followed by amino acids S69 to S208 of KGF2.
225 2555 pEE12.1:TR1.M1-Q193.HSA Amino acids M1 to Q193 of TR1 (including native pEE12.1 1227 1212 1242 Native TR1
signal sequence) fused upstream of mature HSA.
226 2556 pEE12.1:TR1.M1-K194.HSA Amino acids M1 to K194 of TR1 (including native pEE12.1 1228 1213 1243 Native TR1
signal sequence) fused upstream of mature HSA.
227 2557 pEE12.1:TR1.M1-E203.HSA Amino acids M1 to E203 of TR1 (including native pEE12.1 1229 1214 1244 Native TR1
signal sequence) fused upstream of mature HSA.
228 2558 pEE12.1:TR1.M1-Q339.HSA Amino acids M1 to Q339 of TR1 (including native pEE12.1 1230 1215 1245 Native TR1
signal sequence) fused upstream of mature HSA.
229 2571 pC4.OSCAR.R232.HSA M1-R232 of OSCAR fused upstream of mature pC4 1518 1350 1686 1862 1863 Native
HSA. OSCAR
receptor
leader
230 2580 pC4.IFNb(deltaM22, C38S).HSA IFNb fused upstream of mature HSA. The IFNb pC4 1519 1351 1687 Native IFNβ
used in this fusion lacks the first residue of the
mature form of IFNb, which corresponds to M22
of SEQ ID NO: 1687. Also amino acid 38 of SEQ
ID NO: 1687 has been mutated from Cys to Ser.
231 2584 pC4:MPIFsp.KGF2delta28.A63-S208. MPIF signal sequence followed by A63 through pC4 1520 1352 1688 1864 1865 MPIF
HSA S208 of KGF2 followed by mature HSA.
232 2603 pC4:HSA(A14)-EPO(A28-D192. Modified HSA A14 leader fused upstream of pC4 1521 1353 1689 Modified
G140) mature HSA which is fused upstream of A28 HSA (A14)
through D192 of EPO. Amino acid 140 of EPO is
a ‘G’.
233 2604 pC4:HSA(S14)-EPO(A28-D192. Modified HSA S14 leader fused upstream of pC4 1522 1354 1690 Modified
G140) mature HSA which is fused upstream of A28 HSA (S14)
through D192 of EPO. Amino acid 140 of EPO is
a ‘G’.
234 2605 pC4:HSA(G14)-EPO(A28-D192. Modified HSA G14 leader fused upstream of pC4 1523 1355 1691 Modified
G140) mature HSA which is fused upstream of A28 HSA (G14)
through D192 of EPO. Amino acid 140 of EPO is
a ‘G’.
235 2606 pC4:HSA#64.KGF2D28.A63-S208 A63 through S208 of KGF2 fused downstream of pC4 1524 1356 1692 1866 1867 Modified
mature HSA and the modified #64 leader sequence. HSA #64
236 2607 pC4:HSA#65.KGF2D28.A63-S208 A63 through S208 of KGF2 downstream of mature pC4 1525 1357 1693 1868 1869 Modified
HSA and the modified #65 leader sequence. HSA #65
237 2608 pC4:HSA#66.KGF2D28.A63-S208 A63 through S208 of KGF2 fused downstream of pC4 1526 1358 1694 1870 1871 Modified
mature HSA and the modified #66 leader sequence. HSA #66
238 2623 pC4:(AGVSG, 14-18) A modified HSA A14 leader followed by mature pC4 1527 1359 1695 Modified
HSA.HLDOU18.K23-R429 HSA and amino acids K23 through R429 of HSA (A14)
HLDOU18. leader
239 2624 pC4:(SGVSG, 14-18) Modified HSA S14 leader followed by mature pC4 1528 1360 1696 Modified
HSA.HLDOU18.K23-R429 HSA and amino acids K23 to R429 of HLDOU18. HSA (S14)
leader
240 2625 pC4:(GGVSG, 14-18) A modified HSA G14 leader sequence followed by pC4 1529 1361 1697 Modified
HSA.HLDOU18.K23-R429 mature HSA and amino acids K23 through R429 of HSA (G14)
HLDOU18. leader
241 2630 pC4:HSA.KGF2D28.A63-S208#2 Amino acids A63 to S208 of KGF-2 fused to the pC4 1530 1362 1698 1872 1873 HSA
C-terminus of HSA.
242 2631 pEE12.1:(AGVSG, 14-18) A modified HSA A14 leader sequence followed by pEE12.1 1531 1363 1699 Modified
HSA.HLDOU18.K23-R429 mature HSA and amino acids K23 through R429 of HSA (A14)
HLDOU18. leader
243 2632 pEE12.1:(SGVSG, 14-18) Modified HSA S14 leader followed by mature pEE12.1 1532 1364 1700 Modified
HSA.HLDOU18.K23-R429 HSA and amino acids K23 to R429 of HLDOU18. HSA (S14)
leader
244 2633 pEE12.1:(GGVSG, 14-18) A modified HSA G14 leader sequence followed by pEE12.1 1533 1365 1701 Modified
HSA.HLDOU18.K23-R429 mature HSA and amino acids K23 through R429 of HSA (G14)
HLDOU18. leader
245 2637 pSAC35:HSA.GCSF.T31-P207 HSA/kex2 leader fused upstream of mature HSA pScNHSA 1534 1366 1702 1874 1875 HSA/kex2
followed by T31 through P207 of GCSF (SEQ ID
NO: 1702).
246 2638 pPPC007:116A01.HSA scFv I116A01 with C-terminal HSA fusion, where pPPC007 1535 1367 1703 1876 1877 scFvI006A01
the mature form of HSA lacks the first 8 amino
acids.
247 2647 pSAC35:T7.HSA. The T7 peptide (SEQ ID NO: 1704) of Tumstatin pScCHSA 1536 1368 1704 1878 1879 HSA/kex2
was fused with a C-terminal HSA and N terminal
HSA/kex2 leader.
248 2648 pSAC35:T8.HSA The T8 peptide (SEQ ID NO: 1705) of Tumstatin pScCHSA 1537 1369 1705 1880 1881 HSA/kex2
is fused upstream to mature HSA and downstream
from HSA/kex2.
249 2649 pSAC35:HSA.T7 The T7 peptide (SEQ ID NO: 1706) of Tumstatin pScNHSA 1538 1370 1706 1882 1883 HSA/kex2
was fused with a N-terminal HSA/kex2 signal
sequence.
250 2650 pSAC35:HSA.T8 The T8 peptide (SEQ ID NO: 1767) of Tumstatin pScNHSA 1539 1371 1707 1884 1885 HSA/kex2
is fused downstream to HSA/kex2 signal sequence
and mature HSA.
251 2656 pSac35:Insulin(KR.GGG.KR). Synthetic gene coding for a single-chain insulin pScCHSA 1540 1372 1708 1886 1887 HSA/kex2
HSA with HSA at C-terminus. Contains a modified loop
for processing resulting in correctly disulfide
bonded insulin coupled to HSA.
252 2667 pSAC35:HSA.T1249 T1249 fused downstream of full length HSA pSAC35 1178 1179 1180 1181 1182 HSA
253 2668 pSac35:HSA.Insulin(KR.GGG. Synthetic gene coding for insulin with FL HSA at pScNHSA 1541 1373 1709 1888 1889 HSA
KR) N-terminus. Contains a modified loop for
processing resulting in correctly disulfide bonded
insulin coupled to HSA.
254 2669 pSac35:Insulin(GGG.KK).HSA Synthetic gene coding for a single-chain insulin pScCHSA 1542 1374 1710 1890 1891 HSA/kex2
with HSA at C-terminus. Contains a modified
loop.
255 2670 pSAC35:T1249.HSA T1249 fused downstream of HSA/kex2 leader and pSAC35 1183 1179 1180 1184 1185 HSA/kex2
upstream of mature HSA.
256 2671 pSac35:HSA.Insulin(GGG.KK) Synthetic gene coding for a single-chain insulin pScNHSA 1543 1375 1711 1892 1893 HSA
with HSA at N-terminus. Contains a modified
loop for greater stability.
257 2672 pSAC35:HSA.T20 Amino terminus of T20 (codon optimized) fused pSAC35 1186 1187 1188 1189 1190 HSA
downstream of full length HSA
258 2673 pSAC35:T20.HSA Amino terminus of T20 (codon optimized) fused pSAC35 1191 1187 1188 1192 1193 HSA/kex2
downstream of HSA/kex2 leader and upstream of
mature HSA.
259 2700 pSAC35:HSA.GCSF.T31-R199 C-terminal deletion of GCSF fused downstream of pSAC35 1544 1376 1712 1894 1895 HSA/kex2
mature HSA.
260 2701 pSAC35:HSA.GCSF.T31-H200 C-terminal deletion of GCSF fused downstream of pScNHSA 1545 1377 1713 1896 1897 HSA/kex2
mature HSA.
261 2702 pSAC35:HSA.GCSF.T31-L201 HSA/kex2 leader followed by mature HSA and pSAC35 1194 1195 1196 1197 1198 HSA/kex2
amino acids T31-L201 of GCSF (corresponding to
amino acids T1 to L171 of SEQ ID NO: 1196).
262 2703 pSAC35:HSA.GCSF.A36-P204 HSA/kex2 leader followed by mature HSA and pScNHSA 1546 1378 1714 1898 1899 HSA/kex2
amino acids A36-P204 of GCSF.
263 2714 pC4:HSASP.PTH34(2)/HSA PTH34 double tandem repeats fused downstream pC4 1199 1200 1201 1202 1203 HSA leader
of HSA leader (with the KEX site deleted - last 6 minus Kex
amino acids of the leader) and upstream of mature site
HSA.
264 2724 pSAC35.sCNTF.HSA HSA/Kex2 fused to CNTF, and then fused to pSAC35 1547 1379 1715 1900 1901 HSA/kex2
mature HSA.
265 2725 pSAC35:HSA.sCNTF HSA/Kex2 fused to mature HSA and then to CNTF pSAC35 1548 1380 1716 1902 1903 HSA/kex2
266 2726 pSac35.INV.GYGinsulin.HSA Synthetic gene coding for a single-chain insulin pSAC35 1549 1381 1717 1904 1905 Invertase
with HSA at C-terminus. The signal peptide of
invertase is used for this construct.
267 2727 pSac35.INV.GYGinsulin(del Synthetic gene coding for a single-chain insulin pSAC35 1550 1382 1718 1906 1907 invertase
F1).HSA with HSA at C-terminus. Construct uses the
invertase signal peptide and is lacking the first
amino acid (F) of mature human insulin.
268 2749 pEE12.1.OSCAR.R232.HSA Amino acids M1 through R232 of OSCAR fused pEE12.1 1551 1383 1719 1908 1909 Native
upstream of mature HSA. OSCAR
leader
269 2784 pSAC35:Insulin(GYG)-HSA Synthetic gene coding for a single-chain insulin pSAC35 1552 1384 1720 1910 1911 invertase
codon optimized with HSA at C-terminus.
270 2789 pSAC35:Insulin(GGG).HSA Synthetic gene coding for a single-chain insulin pSAC35 1553 1385 1721 1912 1913 invertase
(codon optimized) with HSA at C-terminus.
271 2791 pEE12.1:HSAsp.PTH34(2X). Parathyroid hormone is fused in tandem and pEE12.1 1554 1386 1722 HSA leader
HSA upstream of mature HSA and downstream from minus Kex
HSA signal peptide (with the KEX site deleted - site
last 6 amino acids of the leader)
272 2795 pC4:HSA(A14)-IFNb.M22-N187 The mature form of IFNb is fused to the C- pC4 1555 1387 1723 Modified
terminus of HSA, which contains an modified HSA (A14)
signal peptide, designed to improve processing and
homogeneity.
273 2796 pC4:HSA(S14)-IFNb.M22-N187 The mature form of IFNb is fused to the C- pC4 1556 1388 1724 Modified
terminus of HSA, which contains a modified signal HSA (S14)
peptide, designed to improve processing and
homogeneity.
274 2797 pC4:HSA(G14)-IFNb.M22-N187 The mature form of IFNb is fused to the C- pC4 1557 1389 1725 Modified
terminus of HSA, which contains an modified HSA (G14)
signal peptide.
275 2798 pSAC35:Somatostatin(S14).HSA A 14 amino acid peptide of Somatostatin fused pScCHSA 1558 1390 1726 1914 1915 HSA/kex2
downstream of HSA/kex2 leader and upstream of
mature HSA.
276 2802 pSAC35:GLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is fused pScNHSA 1559 1391 1727 HSA/kex2
36(A8G)).IP2.HSA downstream from the HSA/kex2 leader sequence
and upstream from the intervening peptide-2 of
proglucagon peptide and upstream from mature
HSA.
277 2803 pSAC35:GLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is pScCHSA 1231 1216 1246 1261 1262 HSA/kex2
36(A8G))x2.HSA tandemly repeated and fused downstream of the
HSA/kex2 signal sequence, and upstream of
mature HSA.
278 2804 pSAC35:coGLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is pScCHSA 1232 1217 1247 1263 1264 HSA/kex2
36(A8G))x2.HSA tandemly repeated and fused downstream of the
HSA/kex2 signal sequence, and upstream of
mature HSA.
279 2806 pC4:HSA#65.salmoncalcitonin. Modified HSA leader #65 followed by mature pC4 1560 1392 1728 1916 1917 Modified
C1-G33 HSA and C1-G33 of salmon calcitonin. HSA #65
280 2821 pSac35.delKex2.Insulin(GYG). Synthetic gene coding for a single-chain insulin pScCHSA 1561 1393 1729 Modified
HSA with HSA at C-terminus. The kex2 site has been HSA/kex2,
deleted from the HSA/KEX2 signal peptide. lacking the
Kex2 site.
281 2822 pSac35.alphaMF.Insulin(GYG). Synthetic gene coding for a single-chain insulin pSAC35 1562 1394 1730 1920 1921 MFα-1
HSA with HSA at C-terminus. The signal peptide of
alpha mating factor (MFα-1) is used for this
construct.
282 2825 pSAC35:HSA.Somatostatin(S14) 14 amino acid peptide of Somatostatin was fused pScNHSA 1563 1395 1731 1922 1923 HSA/kex2
downstream of HSA/kex2 leader and mature HSA.
283 2830 pSAC35:S28.HSA 28 amino acids of somatostatin fused downstream pScCHSA 1564 1396 1732 1924 1925 HSA/kex2
of HSA/kex2 leader and upstream of mature HSA.
284 2831 pSAC35:HSA.S28 28 amino acids of somatostatin fused downstream pScNHSA 1565 1397 1733 1926 1927 HSA/kex2
of HSA/kex2 leader and mature HSA.
285 2832 pSAC35:Insulin.HSA (yeast Long-acting insulin peptide fused upstream of pScCHSA 1566 1398 1734 1928 1929 invertase
codon optimized) mature HSA.
286 2837 pSAC35:CKB1.K21-N93: K21-N93 of CKB1 (fragment shown as K2 to N74 pScCHSA 1567 1399 1735 1930 1931 HSA/kex2
HSA of SEQ ID NO: 1735) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
287 2838 pSAC35:CKB1.T22-N93: T22-N93 of CKB1 (fragment shown as T3 to N74 pScCHSA 1568 1400 1736 1932 1933 HSA/kex2
HSA of SEQ ID NO: 1736) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
288 2839 pSAC35:CKB1.E23-N93: E23-N93 of CKB1 (fragment shown as E4 to N74 pScCHSA 1569 1401 1737 1934 1935 HSA/kex2
HSA of SEQ ID NO: 1737) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
289 2840 pSAC35:CKB1.S24-N93: S24-N93 of CKB1 (fragment shown as S5 to N74 pScCHSA 1570 1402 1738 1936 1937 HSA/kex2
HSA of SEQ ID NO: 1738) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
290 2841 pSAC35:CKB1.S25-N93: S25-N93 of CKB1 (fragment shown as S6 to N74 pScCHSA 1571 1403 1739 1938 1939 HSA/kex2
HSA of SEQ ID NO: 1739) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
291 2842 pSAC35:CKB1.S26-N93: S26-N93 of CKB1 (fragment shown as S7 to N74 pScCHSA 1572 1404 1740 1940 1941 HSA/kex2
HSA of SEQ ID NO: 1740) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
292 2843 pSAC35:CKB1.R27-N93: R27-N93 of CKB1 (fragment shown as R8 to N74 pScCHSA 1573 1405 1741 1942 1943 HSA/kex2
HSA of SEQ ID NO: 1741) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
293 2844 pSAC35:CKB1.P29-N93: P29-N93 of CKB1 (fragment shown as P10 to N74 pScCHSA 1574 1406 1742 1944 1945 HSA/kex2
HSA of SEQ ID NO: 1742) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
294 2845 pSAC35:CKB1.Y30-N93: Y30-N93 of CKB1 (fragment shown as Y11 to pScCHSA 1575 1407 1743 1946 1947 HSA/kex2
HSA N74 of SEQ ID NO: 1743) fused downstream of the
HSA/kex2 leader and upstream of mature HSA.
295 2849 pC4.MPIFsp.CKB1.G28-N93. G28-N93 of CKB1 (fragment shown as G9 to N74 pC4 1576 1408 1744 1948 1949 MPIF
HSA of SEQ ID NO: 1744) fused downstream of the
MPIF signal peptide and upstream of mature HSA.
296 2872 pSAC35:HSA.IFNaA(C1-Q91)/ This construct contains a hybrid form of IFNaA pSAC35 1309 1310 1311 1312 1313 HSA/kex2
D(L93-E166) and IFNaD fused downstream of mature HSA.
297 2873 pSAC35:HSA.IFNaA(C1-Q91)/ This construct contains a hybrid form of IFNaA pSAC35 1314 1315 1316 1317 1318 HSA/kex2
B(L93-E166) and IFNaB fused downstream of mature HSA.
298 2874 pSAC35:HSA.IFNaA(C1-Q91)/ This construct contains a hybrid form of IFNaA pSAC35 1319 1320 1321 1322 1323 HSA/kex2
F(L93-E166) and IFNaF fused downstream of mature HSA.
299 2875 pSAC35:HSA.IFNaA(C1Q-62)/ This construct contains a hybrid form of IFNaA pSAC35 1324 1325 1326 1327 1328 HSA/kex2
D(Q64-E166) and IFNaD fused downstream of mature HSA.
300 2876 pSAC35:HSA.IFNaA(C1-Q91)/ This construct contains a hybrid form of IFNaA pSAC35 1329 1330 1331 1332 1333 HSA/kex2
D(L93-E166); and IFNaD fused downstream of mature HSA.
R23K, A113V
301 2877 pSAC35:KT.Insulin.HSA Killer toxin signal peptide fused to synthetic gene pScCHSA 1577 1409 1745 1950 1951 Killer toxin
coding for a single-chain insulin with C-terminal
HSA
302 2878 pSAC35:AP.Insulin.HSA Acid phospatase signal peptide fused to synthetic pSAC35 1578 1410 1746 Acid
gene coding for a single-chain insulin with C- phosphatase
terminal HSA.
303 2882 pSac35.alphaMFprepro.Insulin MFα-1 prepro signal followed by GYG insulin pSAC35 1579 1411 1747 MFα-1
(GYG).HSA followed by mature HSA.
304 2885 pSac35.alphaMFpreproEEA.Insulin Yeast MFα-1 prepro signal followed by GYG pSAC35 1580 1412 1748 Yeast
(GYG).HSA insulin follwed by mature HSA. MFα-1
305 2886 pSAC35:HSA.GCSF.P40-P204 HSA/kex2 signal peptide followed by mature HSA pSAC35 1581 1413 1749 1952 1953 HSA/kex2
followed by GCSF (P40-P204).
306 2887 pSAC35:HSA.GCSF.P40-L201 HSA/kex2 signal peptide followed by mature HSA pSAC35 1582 1414 1750 1954 1955 HSA/kex2
followed by GCSF (P40-L201).
307 2888 pSAC35:HSA.GCSF.Q41-L201 HSA/kex2 signal peptide followed by mature HSA pSAC35 1583 1415 1751 1956 1957 HSA/kex2
followed by GCSF (Q41-L201).
308 2889 pSAC35:HSA.GCSF.Q41-P204 HSA/kex2 signal peptide followed by mature HSA pSAC35 1584 1416 1752 1958 1959 HSA/kex2
followed by GCSF (Q41-P204).
309 2890 pC4.HSA.GCSF.T31-P204 HSA/kex2 signal peptide followed by mature HSA pC4 1585 1417 1753 1960 1961 HSA/kex2
followed by GCSF (T31-P204).
310 2891 pGAP.alphaMF.Insulin(GYG). Synthetic gene coding for a single-chain insulin pYPGaf 1586 1418 1754 1962 1963 HSA/kex2
HSA with HSA at C-terminus. The signal peptide of
HSA/kex2 is used for this construct.
311 2897 pGAP.Insulin(KR.GGG.KR). Long-acting insulin analog using a synthetic gene pYPGaf 1587 1419 1755 1964 1965 HSA/kex2
HSA coding for a single-chain insulin with HSA at C-
terminus. Contains a modified loop for processing
resulting in correctly disulfide bonded insulin
coupled to HSA
312 2900 pSAC:GLP-1(7-36)x2.HSA GLP-1(7-36) is tandemly repeated and then fused pScCHSA 1233 1218 1248 1265 1266 HSA/kex2
downstream of the HSA/kex2 signal sequence and
upstream of mature HSA.
313 2901 pSAC35:IL22.A18-P202. Amino acids A18-P202 of IL22 fused downstream pSAC35 1588 1420 1756 1966 1967 HSA/kex2
HSA of HSA/kex2 leader and upstream of mature HSA.
314 2902 pSAC35: A 14 amino acid peptide of Somatostatin, an pScCHSA 1589 1421 1757 1968 1969 HSA/kex2
Somatostatin(S14(A-G)).HSA inhibitor of growth hormone, synthesized as a C-
terminal HSA fusion. Somatostatin has an alanine
to glycine change at amino acid 1 of SEQ ID
NO: 1757.
315 2903 pSAC35:HSA.A18-P202. Amino acids A18-P202 of IL22 fused downstream pSAC35 1590 1422 1758 1970 1971 HSA
IL22 of full length HSA.
316 2904 pSAC35:GLP-1(9-36).GLP- Amino acids E100 to R127 of preproglucagon pScCHSA 1234 1219 1249 1267 1268 HSA/kex2
1(7-36).HSA (SEQ ID NO: 1249) (hereinafter, this particular
mutant is referred to as GLP-1(9-36)) is fused
downstream from the HSA/kex2 signal sequence
and upstream from GLP-1(7-36), and mature HSA.
317 2908 pSAC35:HSA.HCE1P80 Mature HSA fused downstream of the HSA/kex2 pSAC35 1591 1423 1759 1972 1973 HSA/kex2
leader and upstream of HCE1P80.
318 2909 pSAC35:HSA.HDRMI82 Mature HSA fused downstream of the HSA/kex2 pSAC35 1592 1424 1760 1974 1975 HSA/kex2
leader sequence and upstream of HDRMI82.
319 2910 pSAC35:HSA.RegIV Mature HSA fused downstream of the HSA/kex2 pSAC35 1593 1425 1761 1976 1977 HSA/kex2
leader sequence and upstream of RegIV.
320 2915 pC4:HSA#65.humancalcitonin. Modified HSA leader #65 followed by mature pC4 1594 1426 1762 1978 1979 Modified
C1-G33 HSA and C98 through G130 of SEQ ID NO: 1762. HSA #65
321 2930 pC4.MPIF.Insulin(GYG).HSA Insulin is downstream of an MPIF signal peptide pC4 1595 1427 1763 1980 1981 MPIF
and upstream of mature HSA.
322 2931 pC4.HSA.Insulin(GYG) Synthetic gene coding for a mature single-chain pC4 1596 1428 1764 1982 1983 Modified
insulin fused downstream of the modified HSA HSA (A14)
A14 leader and mature HSA. leader
323 2942 pSac35.TA57.Insulin(GYG). The TA57 Propeptide fused to a single chain pScNHSA 1597 1429 1765 1984 1985 TA57
HSA insulin (GYG), and then mature HSA. propeptide
324 2943 pSAC35:HSA.T7.T7.T74-L98 Dimer construct-HSA/kex2 leader followed by pScNHSA 1598 1430 1766 1986 1987 HSA/kex2
mature HSA followed by two copies of T7 peptide
(SEQ ID NO: 1766) of Tumstatin.
325 2944 pSAC:HSA.T8.T8.K69-S95 HSA/kex2 leader followed by mature HSA pScNHSA 1599 1431 1767 1988 1989 HSA/kex2
followed by two copies of T8 peptide (SEQ ID
NO: 1767) of Tumstatin
326 2945 pSAC35:GLP-1(7- Amino acids H98 to R127 of preproglucagon (SEQ pScCHSA 1235 1220 1250 1269 1270 HSA/kex2
36(A8S)).GLP-1(7-36).HSA ID NO: 1250) is mutated at position 99 from
alanine to serine (hereinafter, this particular mutant
is referred to as GLP-1(7-36(A8S)), which is fused
downstream from the HSA/kex2 signal sequence
and upstream from GLP-1(7-36), and mature HSA.
327 2946 pSAC:T1249(x2).HSA This dimer represents the wild type sequence for pScCHSA 1600 1432 1768 1990 1991 HSA/kex2
T1249. Both dimers have been yeast codon
optimized. The second dimer was optimized to be
different from the first (at the wobble position) to
ensure good amplification. Construct has the
HSA/kex2 leader followed by T1249 dimer
followed by mature HSA.
328 2947 pSAC:CKb-1δ8(x2).HSA Invertase signal peptide followed by amino acids pSAC35 1601 1433 1769 1992 1993 invertase
G28-N93 of full length CKβ1 (SEQ IDNO: 1769),
tandemly repeated, followed by mature HSA.
329 2964 pSAC35:GLP-1(7- GLP-1(7-36) is tandemly repeated as a dimer and pSAC35 1236 1221 1251 1271 1272 HSA/kex2
36)x2.HSA fused downstream from the HSA/kex2 leader
sequence and upstream from mature HSA.
330 2965 pC4:MPIFspP.PTH(1-34). MPIF signal peptide followed by 34 amino acids of pC4 1602 1434 1770 1994 1995 MPIF
HSA PTH followed by mature HSA.
331 2966 pEE12:MPIFsp.PTH(1-34). MPIF signal peptide followed by 34 amino acids PEE12.1 1603 1435 1771 1996 1997 MPIF
HSA of PTH followed by mature HSA.
332 2982 pSAC35:GLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is fused pScCHSA 1237 1222 1252 1273 1274 HSA/kex2
36(A8G).GLP-1(7-36).HSA downstream from the HSA/kex2 signal sequence
and upstream from GLP-1(7-36) and mature HSA.
333 2983 pC4.HSA.Growth Modified (A14) HSA leader followed by mature pC4 1604 1436 1772 1998 1999 Modified
Hormone.F27-F-217 HSA followed by F27 through F217 of growth HSA (A14)
hormone (corresponding to amino acids F1 to F191
of SEQ ID NO: 1772).
334 2986 pSac35.y3SP.TA57PP.Insulin The TA57 Propeptide fused to a single chain pScCHSA 1605 1437 1773 2000 2001 TA57
(GYG).HSA insulin (GYG), and then mature HSA. propeptide
335 3025 pSAC35:INU.Insulin.HSA Inulinase signal peptide is fused upstream of pScCHSA 1606 1438 1774 2002 2003 inulinase
single chain insulin (GYG) and HSA.
336 3027 pSAC35:INV.GLP-1(7- Invertase signal peptide followed by GLP-1(7- pSAC35 1607 1439 1775 2004 2005 invertase
36A8G)x2.HSA 36(A8G)) (SEQ ID NO: 1808) tandemly repeated
as a dimer, followed by mature HSA.
337 3028 pSAC35:INV.GLP-1(7- Invertase signal peptide followed by GLP-1(7- pSAC35 1608 1440 1776 2006 2007 invertase
36(A8G)).GLP-1(7-36).HSA 36(A8G)) (SEQ ID NO: 1808), then GLP-1(7-
36(A8G)), and then mature HSA.
338 3045 pSAC35:DeltaKex.GLP-1(7- HSA/kex2 signal sequence, minus the last six pSAC35 1609 1440 1776 2008 2009 HSA/kex2
36A8G)x2.HSA amino acids of the leader, is fused to GLP-1(7- last six amino
36(A8G)) (SEQ ID NO: 1808) which is tandemly acids
repeated as a dimer, followed by mature HSA.
339 3046 pSAC35:Delta Kex.GLP-1(7- HSA/kex2 signal sequence, minus the last six pSAC35 1610 1440 1776 2010 2011 HSA/kex2
36A8G).GLP-1(7-36).HSA amino acids of the leader, is fused to GLP-1(7- last six amino
36(A8G)) (SEQ ID NO: 1808), GLP-1(7-36), and acids
mature HSA.
340 3047 pSAC35:HSA.Tum5 Full length HSA fused to the Tum5 peptide (SEQ pScNHSA 1611 1443 1779 2012 2013 HSA
ID NO: 1779) of Tumstatin.
341 3048 pSAC35:Tum5.HSA. The Tum5 peptide (SEQ ID NO: 1780) of pScCHSA 1612 1444 1780 2014 2015 HSA/kex2
Tumstatin is fused to HSA and HSA/kex2 leader.
342 3049 pC4.HSA.HCE1P80.D92-L229 Amino acids D92 to L229 of HCE1P80 are fused pC4 1613 1445 1781 2016 2017 HSA
downstream of the full length HSA.
343 3050 pC4.HSA.HCE1P80.A20-L229 Amino acids A20-L229 of HCE1P80 are fused pC4 1614 1446 1782 2018 2019 HSA
downstream of the full length human HSA
344 3051 pSAC35.HSA.HCE1P80.D92-L229 Amino acids D92 to L229 of HCE1P80, a member pSAC35 1615 1447 1783 2020 2021 HSA
of the C1q family of proteins, are fused
downstream of the full length human HSA
345 3052 pSAC35.HSA.HCE1P80.A20-L229 Amino acids A20-L229 of HCE1P80 are fused pSAC35 1616 1448 1784 2022 2023 HSA
downstream of the full length human HSA
346 3053 pC4.HSA.HDALV07.K101-N244 The globular domain of adiponectin (amino acids pC4 1617 1449 1785 2024 2025 HSA
K101-N244) has been inserted downstream of full
length human HSA
347 3055 pSAC35.HSA.HDALV07(GD) Full length HSA followed by amino acids K101-N244 pSAC35 1618 1450 1786 2026 2027 HSA
of HDALV07(GD)/Adiponectin.
348 3056 pSAC35.HSA.HDALV07.MP Full length HSA followed by amino acids Q18 to pSAC35 1619 1451 1787 2028 2029 HSA
N244 of HDALV07.
349 3066 pSAC35:CKB-1d8.GLP-1(7- Invertase signal peptide followed by amino acids pScCHSA 1620 1452 1788 2030 2031 invertase
36).HSA G28-N93 of full length CKβ1 (SEQ IDNO: 1788),
followed by GLP-1(7-36), followed by mature
HSA.
350 3069 pSAC35:INU.GLP-1(7- The inulinase signal sequence is fused to GLP-1(7- pSAC35 1621 1453 1789 2032 2033 inulinase
36(A8G))x2.HSA 36(A8G)) (SEQ ID NO: 1808), which is tandemly
repeated as a dimer and fused to mature HSA.
351 3070 pSAC35:KT.GLP-1(7- GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is pSAC35 1280 1281 1282 1283 1284 Killer toxin
36(A8G))x2.HSA tandemly repeated as a dimer and fused upstream
from mature HSA and downstream from the killer
toxin signal sequence.
352 3071 pSAC35:MAF.GLP-1(7- The yeast mating factor α-1 (hereinafter MFα-1) pSAC35 1622 1454 1790 2034 2035 MFα-1
36(A8G))x2.HSA signal sequence is fused to tandemly repeated
copies of GLP-1(7-36(A8G)) (SEQ ID NO: 1808),
which are fused to mature HSA.
353 3072 pSAC35:AP.GLP-1(7- The acid phosphatase signal sequence is fused to pSAC35 1623 1455 1791 2036 2037 Acid
36(A8G))x2.HSA tandemly repeated copies of GLP-1(7-36(A8G)) phosphatase
(SEQ ID NO: 1808), which are fused to mature
HSA.
354 3085 pSAC35:MAF.GLP-1(7- The yeast mating factor α-1 (hereinafter MFα-1) pSAC35 1624 1456 1792 2038 2039 MFα-1
36(A8G)).GLP-1(7-36).HSA signal sequence is fused to GLP-1(7-36(A8G))
(SEQ ID NO: 1808), GLP-1(7-36), and mature
HSA.
355 3086 pSAC35:INU.GLP-1(7- The inulinase signal sequence is fused to GLP-1(7- pSAC35 1625 1457 1793 2040 2041 inulinase
36(A8G)).GLP-1(7-36).HSA 36(A8G)) (SEQ ID NO: 1808), GLP-1(7-36), and
mature HSA.
356 3087 pSAC35:AP.GLP-1(7- The acid phosphatase signal sequence is fused to pSAC35 1626 1458 1794 2042 2043 Acid
36(A8G)).GLP-1(7-36).HSA GLP-1(7-36(A8G)) (SEQ ID NO: 1808), GLP-1(7- phosphatase
36), and mature HSA.
357 3088 pSAC35.HSA.C-Peptide HSA/kex2 signal peptide, followed by HSA, pSAC35 1627 1459 1795 2044 2045 HSA/kex2
followed by the C-Peptide sequence.
358 3105 pSAC35:INV.t9HCC-1.G28-N93: Invertase signal peptide followed by amino acids pSAC35 1628 1460 1796 2046 2047 invertase
spc.HSA G28 to N93 of HCC-1 fused upstream of a spacer
and mature HSA.
359 3106 pSACHSA.HCBOG68 mature HCBOG68 fused downstream of mature pSAC35 1629 1461 1797 HSA/kex2
HSA and the HSA/kex2 leader sequence.
360 3108 pSAC35HSA.PYY Mature PYY fused downstream of mature HSA pSAC35 1630 1462 1798 HSA/kex2
and the HSA/kex2 leader.
361 3109 pSAC35HSA.PYY3-36 HSA/kex2 leader followed by mature HSA and pSAC35 1631 1463 1799 HSA/kex2
then PYY3-36 (SEQ ID NO: 1799).
362 3117 pC4:PYY3-36/HSA HSA leader followed by PYY3-36 (SEQ ID pC4 1632 1464 1800 2048 2049 HSA
NO: 1800) and mature HSA.
363 3118 pSAC35:PYY3-36/HSA HSA/kex2 leader followed by PYY3-36 (SEQ ID pSAC35 1633 1465 1801 2050 2051 HSA/kex2
NO: 1801) and mature HSA.
364 3119 pSAC35:BNP/HSA HSA/kex2 leader followed by BNP and mature pSAC35 1634 1466 1802 2052 2053 HSA/kex2
HSA.
365 3124 pSAC35:INV.CKB1.P29-N93: Invertase signal peptide followed by amino acids pSAC35 1635 1467 1803 2054 2055 invertase
HSA 29 to 93 of full length ckbetal fused to N-terminus
of HSA.
366 3125 pSAC35:INV.CKb-1.R27-N93: Invertase signal peptide followed by amino acids pSAC35 1636 1468 1804 2056 2057 invertase
HSA 27 to 93 of full length ckbetal fused to N-terminus
of HSA.
367 3133 pSac35.ySP.TA57PP.Insulin Variant TA57 propeptide leader followed by single pSAC35 1637 1469 1805 2058 2059 TA57 variant 1
(GYG).HSA chain insulin, followed by mature HSA.
368 3134 pSac35.ySP.TA57PP + S. Insulin Variant TA57 propeptide leader followed by single pSAC35 1638 1470 1806 2060 2061 TA57 variant 2
(GYG).HSA chain insulin, followed by mature HSA.
369 3139 pSAC35:INV.CKB1.G28-N93. Invertase signal peptide followed by amino acids pSAC35 1639 1471 1807 2062 2063 invertase
DAHK.HSA G28-N93 of full length CKβ1 (see, e.g, SEQ
IDNO: 1788), followed by a 16 amino acid linker
derived from the N-terminus of HSA, followed by
mature HSA.
370 3140 pSAC35:GLP1(mut)DAHK.HSA GLP-1(7-36(A8G)) (SEQ ID NO: 1808) is linked to pSAC35 1640 1472 1808 2064 2065 HSA/kex2
mature HSA by a 16 amino acid linker derived
from the N-terminus of HSA. The HSA/kex2
signal sequence is used.
371 3141 pSAC35:Wnt10b/HSA HSA/kex2 leader followed by amino acids N29 to pSAC35 1641 1473 1809 2066 2067 HSA/kex2
K389 of Wnt10b followed by mature HSA.
372 3142 pSAC35:Wnt11/HSA HSA/kex2 leader followed by mature Wnt11 pSAC35 1642 1474 1810 2068 2069 HSA/kex2
followed by mature HSA.
373 3143 pSAC35:herstatin/HSA HSA/kex2 leader followed by amino acids T23 to pSAC35 1643 1475 1811 2070 2071 HSA/kex2
G419 of herstatin followed by mature HSA.
374 3144 pSAC35:adrenomedullin(27-52)/ HSA/kex2 leader followed by amino acids 27-52 pSAC35 1644 1476 1812 2072 2073 HSA/kex2
HSA of adrenomedullin followed by mature HSA.
375 3149 pSAC35.HSA.C-peptide Full length HSA fused to amino acids E7 to Q37 of pSAC35 1645 1477 1813 2074 2075 HSA
tandem SEQ ID NO: 1813, tandemly repeated.
376 3152 pSAC35:INV.CKB1.Met.R27-N93. Invertase signal peptide followed by a Met, pSAC35 1646 1478 1814 2076 2077 invertase
HSA followed by amino acids R27-N93 of full length
CKβ1, followed by mature HSA.
377 3153 pSAC35:INV.CKB1.Met.S26-N93. Invertase signal peptide followed by a Met, pSAC35 1647 1479 1815 2078 2079 invertase
HSA followed by amino acids S26-N93 of full length
CKβ1, followed by mature HSA.
378 3154 pSAC35:INV.CKB1.Met.S25-N93. Invertase signal peptide followed by a Met, pSAC35 1648 1480 1816 2080 2081 invertase
HSA followed by amino acids S25-N93 of full length
CKβ1, followed by mature HSA.
379 3155 pSAC35:INV.CKB1.Met.G28-N93. Invertase signal peptide followed by a Met, pSAC35 1649 1481 1817 2082 2083 invertase
HSA followed by amino acids G28-N93 of full length
CKβ1, followed by mature HSA.
380 3156 pSAC35:INV.CKB1.Met.P29-N93. Invertase signal peptide followed by a Met, pSAC35 1650 1482 1818 2084 2085 invertase
HSA followed by amino acids P29-N93 of full length
CKβ1, followed by mature HSA.
381 3163 pSAC35:HSA.hGH HSA/kex2 leader fused upstream of mature HSA pSAC35 1303 1304 1305 HSA/kex2
and 191 amino acids of hGH.
382 3165 pSAC35:HSA.IFNa HSA fused upstream of IFNα and downstream of pSAC35 1300 1301 1302 HSA/kex2
also named CID 3165, the HSA/kex2 leader.
pSAC35:HSA.INFα
383 3166 pC4:MPIF1.A22-N93.HSA Amino acids A49 to N120 of MPIF (SEQ ID pC4 1651 1483 1819 2086 2087 MPIF
NO: 1821) is fused downstream of MPIF signal
peptide and upstream of mature HSA.
384 3167 pC4:HSA.MPIF1.D45-N120 Full length HSA followed by amino acids D45 pC4 1652 1484 1820 2088 2089 HSA
through N120 of MPIF.
385 3168 PC4:MPIF-1.HSA Amino acids D45 through N120 of MPIF fused pC4 1653 1485 1821 2090 2091 MPIF
downstream of the MPIF signal sequence and
upstream of mature HSA.
386 3169 pSAC35:KT.CKB1.G28-N93. Killer toxin signal sequence fused upstream of pSAC35 1654 1486 1822 Killer toxin
HSA amino acids G28 through N93 of CKB1 (fragment
shown as amino acids G1 to N66 of SEQ ID
NO: 1822) and mature HSA.
387 3170 pSAC35:KT.HA.CKB1.G28-N93. Killer toxin signal sequence followed by HA pSAC35 1655 1487 1823 Killer toxin
HSA dipeptide and amino acids G28 through N93 of
CKB1 (fragment shown as amino acids G1 to N66
of SEQ ID NO: 1823) and mature HSA.
388 3171 pSAC35:sCNTF(M1-G185): C-terminal deletion of CNTF (amino acids M1 pSAC35 1656 1488 1824 2092 2093 HSA/kex2
HSA through G185), fused upstream of mature HSA and
codon optimized for expression in yeast. HSA/kex2
signal sequence is used.
389 3172 pSAC35:HSA:sCNTF(M1-G185) HSA/kex2 signal sequence followed by mature pSAC35 1657 1489 1825 2094 2095 HSA/kex2
HSA and M1 through G185 of CNTF.
390 3184 pC4:HSA.NOGOR.C27-C309 Full length HSA followed by amino acids C27 to pC4 1658 1490 1826 2096 2097 HSA
C309 of the NOGO receptor.
391 3185 pC4.NOGOR.M1-C309.HSA Amino acids M1-C309 of NOGO receptor fused pC4 1659 1491 1827 2098 2099 Native
upstream of mature HSA. NOGO
receptor
392 3194 pC4:HSA(A14)-EPO(A28-D192. Codon optimized EPO(A28-D192.G140) fused pC4 1660 1492 1828 2100 2101 modified
G140)codon opt downstream of mature HSA with a modified HSA HSA (A14)
(A14) signal sequence.
393 3195 pC4:HSA(S14)-EPO(A28-D192. Codon optimized EPO(A28-D192.G140) fused pC4 1661 1493 1829 2102 2103 modified
G140)codon opt downstream of mature HSA and a modified HSA HSA (S14)
(S14) signal sequence.
394 3196 pC4:HSA(G14)-EPO(A28-D192. Codon optimized EPO(A28-D192.G140) fused pC4 1662 1494 1830 2104 2105 modified
G140)codon opt downstream of mature HSA with a modified (G14) (G14)
HSA signal sequence.
395 3197 pC4.MPIF.Insulin(EAE).HSA A single-chain insulin is downstream of the MPIF pC4 1663 1495 1831 MPIF
signal peptide and upstream of mature human
HSA.
396 3198 pSac35.INV.insulin(EAE).HSA Single-chain insulin is downstream of the invertase pSAC35 1664 1496 1832 invertase
signal peptide and upstream of mature human HSA
397 3202 pSAC35:API.d8CKb1/HSA HSA/kex2 leader followed by amino acids “API” pSAC35 1665 1497 1833 2106 2107 HSA/kex2
followed by d8CKb1 and mature HSA. The
sequence of delta 8 for CKB1 is shown in SEQ ID
NO: 1833.
398 3203 pSAC35:ASL.d8CKb1/HSA HSA/kex2 leader followed by amino acids “ASL” pSAC35 1666 1498 1834 2108 2109 HSA/kex2
followed by d8CKb1 and mature HSA.
399 3204 pSAC35:SPY.d8CKb1/HSA HSA/kex2 leader followed by amino acids “SPY” pSAC35 1667 1499 1835 2110 2111 HSA/kex2
followed by d8CKb1 and mature HSA.
400 3205 pSAC35:MSPY.d8CKb1/HSA HSA/kex2 leader followed by amino acids pSAC35 1668 1500 1836 2112 2113 HSA/kex2
“MSPY” followed by d8CKb1 and mature HSA.
401 3206 pSAC35:CPYSC.d8CKb1/HSA HSA/kex2 leader followed by a five amino acid pSAC35 1669 1501 1837 2114 2115 HSA/kex2
linker followed by d8CKb1 and mature HSA.
402 3207 pSAC35:GPY.d8CKb1/HSA HSA/kex2 leader followed by amino acids “GPY” pSAC35 1670 1502 1838 2116 2117 HSA/kex2
followed by d8CKb1 and mature HSA.
403 3208 pSAC35:defensin alpha Amino acids A65-C94 of defensin alpha 1 fused pSAC35 1285 1286 1287 1288 1289 HSA/kex2
1/HSA downstream of the HSA/kex2 leader and upstream
of mature HSA.
404 3209 pSAC35:defensin alpha Amino acids C66-C94 of defensin alpha 2 fused pSAC35 1290 1291 1292 1293 1294 HSA/kex2
2/HSA downstream of the HSA/kex2 leader and upstream
of mature HSA.
405 3210 pSAC35:defensin alpha Amino acids 65-94 of SEQ ID NO1297, with pSAC35 1295 1296 1297 1298 1299 HSA/kex2
3/HSA A65D and F92I mutations, fused downstream of
the HSA/kex2 leader and upstream of mature HSA.
406 3232 pSAC35:CART/HSA HSA/kex2 leader followed by processed active pSAC35 1671 1503 1839 2118 2119 HSA/kex2
cocaine-amphetamine regulated transcript (CART)
(amino acids V69 through L116) followed by
mature HSA.
407 3238 pSAC35:phosphatonin.HSA Phosphatonin fused upstream of HSA. pSAC35 1306 1307 1308 Native
phosphatonin
408 3270 pSAC35:adipokine/HSA HSA/kex2 leader followed by adipokine followed pSAC35 1672 1504 1840 2120 2121 HSA/kex2
by mature HSA.
409 3272 pSAC35.INV:{D}8CK{b}1(x2)/ CKbeta-1 tandem repeat (x2) fusion to the N- pSAC35 1673 1505 1841 2122 2123 invertase
HSA termal HSA. Under the invertase signal peptide.
410 3274 pSAC35:P1pal-12.HSA P1pal-12 pepducin peptide fused upstream of pSAC35 1334 1335 1336 HSA/kex2
mature HSA, and downstream of the HSA/kex2
leader sequence.
411 3275 pSAC35:P4pal-10.HSA P4pal-10 pepducin peptide fused upstream of pSAC35 1337 1338 1339 HSA/kex2
mature HSA, and downstream of the HSA/kex2
leader sequence.
412 3281 pSAC35.PY3-36(x2)/HSA PYY3-36 tandem repeat (x2) fused upstream of pSAC35 1674 1506 1842 2124 2125 HSA/kex2
HSA and downstream of the HSA/kex2 signal
peptide.
413 3282 pSAC35:HSA/PYY3-36(x2) PYY3-36 tandem repeat (x2) fused downstream of pSAC35 1675 1507 1843 2126 2127 HSA/kex2
mature HSA and HSA/kex2 leader.
414 3298 pSAC35:IL21/HSA Amino acids Q30-S162 of IL-21 fused upstream of pSAC35 2167 2157 2177 2188 2189 HSA/Kex2
mature HSA and downstream of HSA/kex2 leader
415 3307 pSAC35:IL4/HSA Amino acids H25-S153 of IL-4 fused upstream of pSAC35 2168 2158 2178 2190 2191 HSA/Kex2
mature HSA and downstream of HSA/kex2 leader
416 3309 pSAC:KT.GLP-1(7- Killer toxin leader sequence followed by GLP-1(7-36 pSAC35 2170 2160 2180 2194 2195 Killer toxin
36(A8G))x2.MSA.E25-A608 (A8G) followed by mature mouse serum
albumin.
417 3312 pSAC35:hOCIL/HSA HSA/kex2 leader followed by amino acids N20 to pSAC35 2171 2161 2181 2196 2197 HSA/Kex2
V149 of hOCIL followed by mature HSA
418 7777 T20:HSA T20 fused downstream of full length HSA pC4 1170 1171 1172 HSA
419 8888 pC4:BNP.HSA Human B-type natriuretic peptide fused upstream pC4 1275 1276 1277 1278 1279 Native BNP
of mature HSA.
420 9999 T1249:HSA T1249 fused downstream of full length HSA pC4 1173 1174 1175 HSA

Table 2 provides a non-exhaustive list of polynucleotides of the invention comprising, or alternatively consisting of, nucleic acid molecules encoding an albumin fusion protein. The first column, “Fusion No.” gives a fusion number to each polynucleotide. Column 2, “Construct ID” provides a unique numerical identifier for each polynucleotide of the invention. The Construct IDs may be used to identify polynucleotides which encode albumin fusion proteins comprising, or alternatively consisting of, a Therapeutic protein portion corresponding to a given Therapeutic Protein:X listed in the corresponding row of Table 1 wherein that Construct ID is listed in column 5. The “Construct Name” column (column 3) provides the name of a given albumin fusion construct or polynucleotide.

The fourth column in Table 2, “Description” provides a general description of a given albumin fusion construct, and the fifth column, “Expression Vector” lists the vector into which a polynucleotide comprising, or alternatively consisting of, a nucleic acid molecule encoding a given albumin fusion protein was cloned. Vectors are known in the art, and are available commercially or described elsewhere. For example, as described in the Examples, an “expression cassette” comprising, or alternatively consisting of, one or more of (1) a polynucleotide encoding a given albumin fusion protein, (2) a leader sequence, (3) a promoter region, and (4) a transcriptional terminator, may be assembled in a convenient cloning vector and subsequently be moved into an alternative vector, such as, for example, an expression vector including, for example, a yeast expression vector or a mammalian expression vector. In one embodiment, for expression in S. cervisiae, an expression cassette comprising, or alternatively consisting of, a nucleic acid molecule encoding an albumin fusion protein is cloned into pSAC35. In another embodiment, for expression in CHO cells, an expression cassette comprising, or alternatively consisting of, a nucleic acid molecule encoding an albumin fusion protein is cloned into pC4. In a further embodiment, a polynucleotide comprising or alternatively consisting of a nucleic acid molecule encoding the Therapeutic protein portion of an albumin fusion protein is cloned into pC4:HSA. In a still further embodiment, for expression in NSO cells, an expression cassette comprising, or alternatively consisting of, a nucleic acid molecule encoding an albumin fusion protein is cloned into pEE12. Other useful cloning and/or expression vectors will be known to the skilled artisan and are within the scope of the invention.

Column 6, “SEQ ID NO:Y,” provides the full length amino acid sequence of the albumin fusion protein of the invention. In most instances, SEQ ID NO:Y shows the unprocessed form of the albumin fusion protein encoded—in other words, SEQ ID NO:Y shows the signal sequence, a HSA portion, and a therapeutic portion all encoded by the particular construct. Specifically contemplated by the present invention are all polynucleotides that encode SEQ ID NO:Y. When these polynucleotides are used to express the encoded protein from a cell, the cell's natural secretion and processing steps produces a protein that lacks the signal sequence listed in columns 4 and/or 11 of Table 2. The specific amino acid sequence of the listed signal sequence is shown later in the specification or is well known in the art. Thus, most preferred embodiments of the present invention include the albumin fusion protein produced by a cell (which would lack the leader sequence shown in columns 4 and/or 11 of Table 2). Also most preferred are polypeptides comprising SEQ ID NO:Y without the specific leader sequence listed in columns 4 and/or 11 of Table 2. Compositions comprising these two preferred embodiments, including pharmaceutical compositions, are also preferred. Moreover, it is well within the ability of the skilled artisan to replace the signal sequence listed in columns 4 and/or 11 of Table 2 with a different signal sequence, such as those described later in the specification to facilitate secretion of the processed albumin fusion protein.

The seventh column, “SEQ ID NO:X,” provides the parent nucleic acid sequence from which a polynucleotide encoding a Therapeutic protein portion of a given albumin fusion protein may be derived. In one embodiment, the parent nucleic acid sequence from which a polynucleotide encoding a Therapeutic protein portion of an albumin fusion protein may be derived comprises the wild type gene sequence encoding a Therapeutic protein shown in Table 1. In an alternative embodiment, the parent nucleic acid sequence from which a polynucleotide encoding a Therapeutic protein portion of an albumin fusion protein may be derived comprises a variant or derivative of a wild type gene sequence encoding a Therapeutic protein shown in Table 1, such as, for example, a synthetic codon optimized variant of a wild type gene sequence encoding a Therapeutic protein.

The eighth column, “SEQ ID NO:Z,” provides a predicted translation of the parent nucleic acid sequence (SEQ ID NO:X). This parent sequence can be a full length parent protein used to derive the particular construct, the mature portion of a patent protein, a variant or fragment of a wildtype protein, or an artificial sequence that can be used to create the described construct. One of skill in the art can use this amino acid sequence shown in SEQ ID NO:Z to determine which amino acid residues of an albumin fusion protein encoded by a given construct are provided by the therapeutic protein. Moreover, it is well within the ability of the skilled artisan to use the sequence shown as SEQ ID NO:Z to derive the construct described in the same row. For example, if SEQ ID NO:Z corresponds to a full length protein, but only a portion of that protein is used to generate the specific CID, it is within the skill of the art to rely on molecular biology techniques, such as PCR, to amplify the specific fragment and clone it into the appropriate vector.

Amplification primers provided in columns 9 and 10, “SEQ ID NO:A” and “SEQ ID NO:B” respectively, are exemplary primers used to generate a polynucleotide comprising or alternatively consisting of a nucleic acid molecule encoding the Therapeutic protein portion of a given albumin fusion protein. In one embodiment of the invention, oligonucleotide primers having the sequences shown in columns 9 and/or 10 (SEQ ID NOS:A and/or B) are used to PCR amplify a polynucleotide encoding the Therapeutic protein portion of an albumin fusion protein using a nucleic acid molecule comprising or alternatively consisting of the nucleotide sequence provided in column 7 (SEQ ID NO:X) of the corresponding row as the template DNA. PCR methods are well-established in the art. Additional useful primer sequences could readily be envisioned and utilized by those of ordinary skill in the art.

In an alternative embodiment, oligonucleotide primers may be used in overlapping PCR reactions to generate mutations within a template DNA sequence. PCR methods are known in the art.

As shown in Table 3, certain albumin fusion constructs disclosed in this application have been deposited with the ATCC®.

TABLE 3
Construct ID Construct Name ATCC Deposit No./Date
1642 pSAC35:GCSF.T31-P204.HSA PTA-3767
Oct. 5, 2001
1643 pSAC35:HSA.GCSF.T31-P204 PTA-3766
Oct. 5, 2001
1812 pSAC35:IL2.A21-T153.HSA PTA-3759
Oct. 4, 2001
1941 pC4:HSA/PTH84(junctioned) PTA-3761
Oct. 4, 2001
1949 pC4:PTH.S1-Q84/HSA (junctioned) PTA-3762
Oct. 4, 2001
1966 pC4:EPO.M1-D192.HSA PTA-3771
also named pC4:EPOM1-D192.HSA Oct. 5, 2001
1981 pC4.HSA-EPO.A28-D192 PTA-3770
Oct. 5, 2001
1997 pEE12.1:EPOM1-D192.HSA PTA-3768
Oct. 5, 2001
2030 pSAC35.ycoIL-2.A21-T153.HSA PTA-3757
Oct. 4, 2001
2031 pSAC35.HSA.ycoIL-2.A21-T153 PTA-3758
Oct. 4, 2001
2053 pEE12:IFNb-HSA PTA-3764
also named pEE12.1:IFNβ-HSA Oct. 4, 2001
2054 pEE12:HSA-IFNb PTA-3941
Dec. 19, 2001
2249 pSAC35:IFNa2-HSA PTA-3763
also named pSAC23:IFNα2-HSA Oct. 4, 2001
2250 pSAC35:HSA.INSULIN(GYG) PTA-3916
also named pSAC35.HSA.INSULING(GYG).F1-N62 Dec. 07, 2001
2255 pSAC35:INSULIN(GYG).HSA PTA-3917
also named pSAC35.INSULING(GYG).F1-N62.HSA Dec. 07, 2001
2276 pSAC35:HSA.INSULIN(GGG) PTA-3918
also named pSAC35.HSA.INSULING(GGG).F1-N58 Dec. 07, 2001
2298 pEE12.1:EPO.R140G.HSA PTA-3760
Oct. 4, 2001
2294 pC4:EPO.R140G.HSA PTA-3742
also named pC4.EPO.R1406.HSA Sept. 28, 2001
2325 pC4.EPO:M1-D192.HSA.Codon opt. PTA-3773
Oct. 5, 2001
2343 pSAC35.INV-IFNA2.HSA PTA-3940
Dec. 19, 2001
2363 pC4.GCSF.HSA.EPO.A28-D192 PTA-3740
Sept. 28, 2001
2373 pC4.GCSF.HSA.EPO.A28-D192.R140G PTA-3741
Sept. 28, 2001
2381 pC4:HSA-IFNa2(C17-E181) PTA-3942
Dec. 19, 2001
2382 pC4:IFNa2-HSA PTA-3939
Dec. 19, 2001
2387 pC4:EPO(G140)-HSA-GCSF.T31-P204 PTA-3919
Dec. 11, 2001
2414 pC4.EPO:M1-D192copt.HSA.GCSF.T31-P204 PTA-3924
also named Dec. 12, 2001
pC4.EPO:M1-D192copt.HAS.GCSF.T31-P204
2441 pEE12.EPO:M1-D192copt.HSA.GCSF.T31-P204 PTA-3923
also named: Dec. 12, 2001
pEE12.EPO:M1-D192copt.HAS.GCSF.T31-P204
2492 pC4.IFNb(deltaM22).HSA PTA-3943
Dec. 19, 2001
3070 pSAC35:KT.GLP-1(7-36(A8G))x2.HSA PTA-4671
Sept. 16, 2002
3165 pSAC35:HSA.IFNa PTA-4670
also named CID 3165, pSAC35:HSA.INFα Sept. 16, 2002
3163 pSAC35:HSA.hGH PTA-4770
Oct. 22, 2002

It is possible to retrieve a given albumin fusion construct from the deposit by techniques known in the art and described elsewhere herein (see, Example 40). The ATCC is located at 10801 University Boulevard, Manassas, Va., 20110-2209, USA. The ATCC deposits were made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.

In a further embodiment of the invention, an “expression cassette” comprising, or alternatively consisting of one or more of (1) a polynucleotide encoding a given albumin fusion protein, (2) a leader sequence, (3) a promoter region, and (4) a transcriptional terminator can be moved or “subcloned” from one vector into another. Fragments to be subcloned may be generated by methods well known in the art, such as, for example, PCR amplification (e.g., using oligonucleotide primers having the sequence shown in SEQ ID NO:A or B), and/or restriction enzyme digestion.

In preferred embodiments, the albumin fusion proteins of the invention are capable of a therapeutic activity and/or biologic activity corresponding to the therapeutic activity and/or biologic activity of the Therapeutic protein corresponding to the Therapeutic protein portion of the albumin fusion protein listed in the corresponding row of Table 1. In further preferred embodiments, the therapeutically active protein portions of the albumin fusion proteins of the invention are fragments or variants of the protein encoded by the sequence shown in SEQ ID NO:X column of Table 2, and are capable of the therapeutic activity and/or biologic activity of the corresponding Therapeutic protein.

Non-Human Albumin Fusion Proteins of Growth Hormone.

In one embodiment, the albumin fusion proteins of the invention comprise one or more Serum Albumin proteins of a non-human animal species, fused in tandem and in-frame either at the N-terminus or the C-terminus to one or more Growth Hormone proteins of the same non-human animal species. Non-human Serum Albumin and Growth Hormone proteins are well known in the art and available in public databases. For example, Table 4 presents accession numbers corresponding to non-human Serum Albumin sequences (column 2) and non-human Growth Hormone sequences (column 3) found in GenBank. In a preferred embodiment, a Serum Albumin protein from a non-human animal species listed in Table 4 is fused to a Growth Hormone protein from the same non-human animal species.

In a specific embodiment, the albumin fusion protein of the invention comprises one or more Bos taurus Serum Albumin proteins listed in Table 4, column 2, fused in tandem and in-frame either at the N-terminus or the C-terminus to one or more Bos taurus Growth Hormone proteins listed in Table 4, column 3.

Fusion proteins comprising fragments or variants of non-human Serum Albumin, such as, for example, the mature form of Serum Albumin, are also encompassed by the invention. Fusion proteins comprising fragments or variants of non-human Growth Hormone proteins, such as, for example, the mature form of Growth Hormone, are also encompassed by the invention. Preferably the non-human Growth Hormone fragments and variants retain growth hormone activity.

Polynucleotides of the invention comprise, or alternatively consist of, one or more nucleic acid molecules encoding a non-human albumin fusion protein described above. For example, the polynucleotides can comprise, or alternatively consist of, one or more nucleic acid molecules that encode a Serum Albumin protein from a non-human animal species listed in Table 4, column 1 (such as, for example, the non-human Serum Albumin reference sequences listed in Table 4, column 2) fused in tandem and in-frame either 5′ or 3′ to a polynucleotide that comprises, or alternatively consists of, one or more nucleic acid molecules encoding the non-human Growth Hormone protein of the corresponding non-human animal species (for example, the Growth Hormone reference sequences listed in Table 4, column 3).

The above-described non-human albumin fusion proteins are encompassed by the invention, as are host cells and vectors containing these polynucleotides. In one embodiment, a non-human albumin fusion protein encoded by a polynucleotide as described above has extended shelf life. In an additional embodiment, a non-human albumin fusion protein encoded by a polynucleotide described above has a longer serum half-life and/or more stabilized activity in solution (or in a pharmaceutical composition) in vitro and/or in vivo than the corresponding unfused Growth Hormone molecule.

The present invention also encompasses methods of preventing, treating, or ameliorating a disease or disorder in a non-human animal species. In certain embodiments, the present invention encompasses a method of treating a veterinary disease or disorder comprising administering to a non-human animal species in which such treatment, prevention or amelioration is desired an albumin fusion protein of the invention that comprises a Growth Hormone portion corresponding to a Growth Hormone protein (or fragment or variant thereof) in an amount effective to treat, prevent or ameliorate the disease or disorder. Veterinary diseases and/or disorders which may be treated, prevented, or ameliorated include growth disorders (such as, for example, pituitary dwarfism), shin soreness, obesity, growth hormone-responsive dermatosis, dilated cardiomyopathy, eating disorders, reproductive disorders, and endocrine disorders.

Non-human albumin fusion proteins of the invention may also be used to promote healing of skin wounds, corneal injuries, bone fractures, and injuries of joints, tendons, or ligaments.

Non-human albumin fusion proteins of the invention may also be used to increase milk production in lactating animals. In a preferred embodiment, the lactating animal is a dairy cow.

Non-human albumin fusion proteins of the invention may also be used to improve body condition in aged animals.

Non-human albumin fusion proteins of the invention may also be used to increase fertility, pregnancy rates, and reproductive success in domesticated animals.

Non-human albumin fusion proteins of the invention may also be used to improve the lean-to-fat ratio in animals raised for consumption, as well as to improve appetite, and increase body size and growth rate.

TABLE 4
Non-Human Serum Albumin Reference
Non-Human Sequence(s): GenBank Protein Non-Human Growth Hormone Reference
Species Accession Nos. Sequence(s): GenBank Protein Accession Nos.
Bos taurus ABBOS, CAA76847, P02769, STBO, BAA06379, A29864, AAF28806,
CAA41735, 229552, AAA51411 AAF28805, AAF28804, P01246, AAF03132,
AAC63901, AAB92549, A36506, I45901, JC1316,
CAA23445, CAA00787, CAA00598, AAA30547,
AAA30546, AAA30545, AAA30544, AAA30543,
AAA30542
Sus scrofa P08835, CAA30970, AAA30988 STPG, PC1017, AAB29947, AAB84359, I46585,
I46584, PC1063, A01516, AAB17619, 226829,
225740, CAA37411, CAA00592, AAA73478,
AAA73477, CAA00356, AAA31046, AAA31045,
AAA31044, AA30543
Equus caballus ABHOS, AAG40944, P35747, STHO, P01245, AAD25992, 227704, AAA21027
CAA52194
Ovis aries ABSHS, P14639, CAA34903 STSH, AAB24467, AAC48679, 228487, 223932,
CAA34098, CAA31063, CAA00828, AAA31527
Salmo salar ABONS2, ABONS2, CAA36643, STONC, P07064, Q07221, P48096, P10814,
CAA43187 P10607, I51186, S03709, JS0179, A23154,
S06489, CAA42431, AAB29165, AAB24612,
Q91221, Q91222, CAA43942, CAA32481,
738042, 224555, CAA00427, AAA50757,
AAA49558, AAA49555, AAA49553, AAA49401,
AAA49406, AAA49403, AAA49402
Gallus gallus ABCHS, P19121, CAA43098 BAB62262, BAB69037, AAK95643, A60509,
AAG01029, BAA01365, P08998, 226895,
CAA31127, CAA35619, AAA48180
Felis catus P49064, S57632, CAA59279, JC4660 JC4632, P46404, AAC00073, AAA96142,
AAA67294
Canis familiaris P49822, S29749, CAB64867, P33711, I46145, AAF89582, AAF21502,
CAA76841, AAB30434 AAD43366, S35790, AAB34229, CAA80601

Polypeptide and Polynucleotide Fragments and Variants

Fragments

The present invention is further directed to fragments of the Therapeutic proteins described in Table 1, albumin proteins, and/or albumin fusion proteins of the invention.

The present invention is also directed to polynucleotides encoding fragments of the Therapeutic proteins described in Table 1, albumin proteins, and/or albumin fusion proteins of the invention.

Even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the Therapeutic protein, albumin protein, and/or albumin fusion protein of the invention, other Therapeutic activities and/or functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) may still be retained. For example, the ability of polypeptides with N-terminal deletions to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptides generally will be retained when less than the majority of the residues of the complete polypeptide are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a mutein with a large number of deleted N-terminal amino acid residues may retain some biological or immunogenic activities. In fact, peptides composed of as few as six amino acid residues may often evoke an immune response.

Accordingly, fragments of a Therapeutic protein corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention, include the full length protein as well as polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the reference polypeptide (i.e., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2). In particular, N-terminal deletions may be described by the general formula m to q, where q is a whole integer representing the total number of amino acid residues in a reference polypeptide (e.g., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein of the invention, or a Therapeutic protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2), and m is defined as any integer ranging from 2 to q minus 6. Polynucleotides encoding these polypeptides are also encompassed by the invention.

In addition, fragments of serum albumin polypeptides corresponding to an albumin protein portion of an albumin fusion protein of the invention, include the full length protein as well as polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the reference polypeptide (i.e., serum albumin, or a serum albumin portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2). In preferred embodiments, N-terminal deletions may be described by the general formula m to 585, where 585 is a whole integer representing the total number of amino acid residues in mature human serum albumin (SEQ ID NO:1038), and m is defined as any integer ranging from 2 to 579. Polynucleotides encoding these polypeptides are also encompassed by the invention. In additional embodiments, N-terminal deletions may be described by the general formula m to 609, where 609 is a whole integer representing the total number of amino acid residues in full length human serum albumin (SEQ ID NO:1094), and m is defined as any integer ranging from 2 to 603. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Moreover, fragments of albumin fusion proteins of the invention, include the full length albumin fusion protein as well as polypeptides having one or more residues deleted from the amino terminus of the albumin fusion protein (e.g., an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2; or an albumin fusion protein having the amino acid sequence disclosed in column 6 of Table 2). In particular, N-terminal deletions may be described by the general formula m to q, where q is a whole integer representing the total number of amino acid residues in the albumin fusion protein, and m is defined as any integer ranging from 2 to q minus 6. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Also as mentioned above, even if deletion of one or more amino acids from the N-terminus or C-terminus of a reference polypeptide (e.g., a Therapeutic protein; serum albumin protein; or albumin fusion protein of the invention) results in modification or loss of one or more biological functions of the protein, other functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) and/or Therapeutic activities may still be retained. For example the ability of polypeptides with C-terminal deletions to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptide generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the C-terminus. Whether a particular polypeptide lacking the N-terminal and/or C-terminal residues of a reference polypeptide retains Therapeutic activity can readily be determined by routine methods described herein and/or otherwise known in the art.

The present invention further provides polypeptides having one or more residues deleted from the carboxy terminus of the amino acid sequence of a Therapeutic protein corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention (e.g., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2). In particular, C-terminal deletions may be described by the general formula 1 to n, where n is any whole integer ranging from 6 to q minus 1, and where q is a whole integer representing the total number of amino acid residues in a reference polypeptide (e.g., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2). Polynucleotides encoding these polypeptides are also encompassed by the invention.

In addition, the present invention provides polypeptides having one or more residues deleted from the carboxy terminus of the amino acid sequence of an albumin protein corresponding to an albumin protein portion of an albumin fusion protein of the invention (e.g., serum albumin or an albumin protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 2). In particular, C-terminal deletions may be described by the general formula 1 to n, where n is any whole integer ranging from 6 to 584, where 584 is the whole integer representing the total number of amino acid residues in mature human serum albumin (SEQ ID NO:1038) minus 1. Polynucleotides encoding these polypeptides are also encompassed by the invention. In particular, C-terminal deletions may be described by the general formula 1 to n, where n is any whole integer ranging from 6 to 608, where 608 is the whole integer representing the total number of amino acid residues in serum albumin (SEQ ID NO:1094) minus 1. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Moreover, the present invention provides polypeptides having one or more residues deleted from the carboxy terminus of an albumin fusion protein of the invention. In particular, C-terminal deletions may be described by the general formula 1 to n, where n is any whole integer ranging from 6 to q minus 1, and where q is a whole integer representing the total number of amino acid residues in an albumin fusion protein of the invention. Polynucleotides encoding these polypeptides are also encompassed by the invention.

In addition, any of the above described N- or C-terminal deletions can be combined to produce a N- and C-terminal deleted reference polypeptide. The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini, which may be described generally as having residues m to n of a reference polypeptide (e.g., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein of the invention, or a Therapeutic protein portion encoded by a polynucleotide or albumin fusion construct described in Table 2, or serum albumin (e.g., SEQ ID NO:1038), or an albumin protein portion of an albumin fusion protein of the invention, or an albumin protein portion encoded by a polynucleotide or albumin fusion construct described in Table 2, or an albumin fusion protein, or an albumin fusion protein encoded by a polynucleotide or albumin fusion construct of the invention) where n and m are integers as described above. Polynucleotides encoding these polypeptides are also encompassed by the invention.

The present application is also directed to proteins containing polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a reference polypeptide sequence (e.g., a Therapeutic protein referred to in Table 1, or a Therapeutic protein portion of an albumin fusion protein of the invention, or a Therapeutic protein portion encoded by a polynucleotide or albumin fusion construct described in Table 2, or serum albumin (e.g., SEQ ID NO: 1038), or an albumin protein portion of an albumin fusion protein of the invention, or an albumin protein portion encoded by a polynucleotide or albumin fusion construct described in Table 2, or an albumin fusion protein, or an albumin fusion protein encoded by a polynucleotide or albumin fusion construct of the invention) set forth herein, or fragments thereof. In preferred embodiments, the application is directed to proteins comprising polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to reference polypeptides having the amino acid sequence of N- and C-terminal deletions as described above. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Preferred polypeptide fragments of the invention are fragments comprising, or alternatively, consisting of, an amino acid sequence that displays a Therapeutic activity and/or functional activity (e.g. biological activity) of the polypeptide sequence of the Therapeutic protein or serum albumin protein of which the amino acid sequence is a fragment.

Other preferred polypeptide fragments are biologically active fragments. Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the polypeptide of the present invention. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity.

Variants

“Variant” refers to a polynucleotide or nucleic acid differing from a reference nucleic acid or polypeptide, but retaining essential properties thereof. Generally, variants are overall closely similar, and, in many regions, identical to the reference nucleic acid or polypeptide.

As used herein, “variant”, refers to a Therapeutic protein portion of an albumin fusion protein of the invention, albumin portion of an albumin fusion protein of the invention, or albumin fusion protein of the invention differing in sequence from a Therapeutic protein (e.g. see “therapeutic” column of Table 1), albumin protein, and/or albumin fusion protein, respectively, but retaining at least one functional and/or therapeutic property thereof as described elsewhere herein or otherwise known in the art. Generally, variants are overall very similar, and, in many regions, identical to the amino acid sequence of the Therapeutic protein corresponding to a Therapeutic protein portion of an albumin fusion protein, albumin protein corresponding to an albumin protein portion of an albumin fusion protein, and/or albumin fusion protein. Nucleic acids encoding these variants are also encompassed by the invention.

The present invention is also directed to proteins which comprise, or alternatively consist of, an amino acid sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, the amino acid sequence of a Therapeutic protein corresponding to a Therapeutic protein portion of an albumin fusion protein of the invention (e.g., the amino acid sequence of a Therapeutic protein:X disclosed in Table 1; or the amino acid sequence of a Therapeutic protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 1 and 2, or fragments or variants thereof), albumin proteins corresponding to an albumin protein portion of an albumin fusion protein of the invention (e.g., the amino acid sequence of an albumin protein portion of an albumin fusion protein encoded by a polynucleotide or albumin fusion construct described in Table 1 and 2; the amino acid sequence shown in SEQ ID NO: 1038; or fragments or variants thereof), and/or albumin fusion proteins. Fragments of these polypeptides are also provided (e.g., those fragments described herein). Further polypeptides encompassed by the invention are polypeptides encoded by polynucleotides which hybridize to the complement of a nucleic acid molecule encoding an albumin fusion protein of the invention under stringent hybridization conditions (e.g., hybridization to filter bound DNA in 6× Sodium chloride/Sodium citrate (SSC) at about 45 degrees Celsius, followed by one or more washes in 0.2×SSC, 0.1% SDS at about 50-65 degrees Celsius), under highly stringent conditions (e.g., hybridization to filter bound DNA in 6× sodium chloride/Sodium citrate (SSC) at about 45 degrees Celsius, followed by one or more washes in 0.1×SSC, 0.2% SDS at about 68 degrees Celsius), or under other stringent hybridization conditions which are known to those of skill in the art (see, for example, Ausubel, F. M. et al., eds., 1989 Current protocol in Molecular Biology, Green publishing associates, Inc., and John Wiley & Sons Inc., New York, at pages 6.3.1-6.3.6 and 2.10.3). Polynucleotides encoding these polypeptides are also encompassed by the invention.

By a polypeptide having an amino acid sequence at least, for example, 95% “identical” to a query amino acid sequence, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a query amino acid sequence, up to 5% of the amino acid residues in the subject sequence may be inserted, deleted, or substituted with another amino acid. These alterations of the reference sequence may occur at the amino- or carboxy-terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.

As a practical matter, whether any particular polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the amino acid sequence of an albumin fusion protein of the invention or a fragment thereof (such as a Therapeutic protein portion of the albumin fusion protein or an albumin portion of the albumin fusion protein), can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci.6:237-245 (1990)). In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is expressed as percent identity. Preferred parameters used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of the subject amino acid sequence, whichever is shorter.

If the subject sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for N- and C-terminal truncations of the subject sequence when calculating global percent identity. For subject sequences truncated at the N- and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are N- and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence. Whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of the present invention. Only residues to the N- and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query residue positions outside the farthest N- and C-terminal residues of the subject sequence.

For example, a 90 amino acid residue subject sequence is aligned with a 100 residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence. This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to made for the purposes of the present invention.

The variant will usually have at least 75% (preferably at least about 80%, 90%, 95% or 99%) sequence identity with a length of normal HA or Therapeutic protein which is the same length as the variant. Homology or identity at the nucleotide or amino acid sequence level is determined by BLAST (Basic Local Alignment Search Tool) analysis using the algorithm employed by the programs blastp, blastn, blastx, tblastn and tblastx (Karlin et al., Proc. Natl. Acad. Sci. USA 87: 2264-2268 (1990) and Altschul, J. Mol. Evol. 36: 290-300 (1993), fully incorporated by reference) which are tailored for sequence similarity searching.

The approach used by the BLAST program is to first consider similar segments between a query sequence and a database sequence, then to evaluate the statistical significance of all matches that are identified and finally to summarize only those matches which satisfy a preselected threshold of significance. For a discussion of basic issues in similarity searching of sequence databases, see Altschul et al., (Nature Genetics 6: 119-129 (1994)) which is fully incorporated by reference. The search parameters for histogram, descriptions, alignments, expect (i.e., the statistical significance threshold for reporting matches against database sequences), cutoff, matrix and filter are at the default settings. The default scoring matrix used by blastp, blastx, tblastn, and tblastx is the BLOSUM62 matrix (Henikoff et al., Proc. Natl. Acad. Sci. USA 89: 10915-10919 (1992), fully incorporated by reference). For blastn, the scoring matrix is set by the ratios of M (i.e., the reward score for a pair of matching residues) to N (i.e., the penalty score for mismatching residues), wherein the default values for M and N are 5 and −4, respectively. Four blastn parameters may be adjusted as follows: Q=10 (gap creation penalty); R=10 (gap extension penalty); wink=1 (generates word hits at every winkth position along the query); and gapw=16 (sets the window width within which gapped alignments are generated). The equivalent Blastp parameter settings were Q=9; R=2; wink=1; and gapw=32. A Bestfit comparison between sequences, available in the GCG package version 10.0, uses DNA parameters GAP=50 (gap creation penalty) and LEN=3 (gap extension penalty) and the equivalent settings in protein comparisons are GAP=8 and LEN=2.

The polynucleotide variants of the invention may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, polypeptide variants in which less than 50, less than 40, less than 30, less than 20, less than 10, or 5-50, 5-25, 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred. Polynucleotide variants can be produced for a variety of reasons, e.g., to optimize codon expression for a particular host (change codons in the human mRNA to those preferred by a bacterial host, such as, yeast or E. coli).

In a preferred embodiment, a polynucleotide of the invention which encodes the albumin portion of an albumin fusion protein is optimized for expression in yeast or mammalian cells. In a further preferred embodiment, a polynucleotide of the invention which encodes the Therapeutic protein portion of an albumin fusion protein is optimized for expression in yeast or mammalian cells. In a still further preferred embodiment, a polynucleotide encoding an albumin fusion protein of the invention is optimized for expression in yeast or mammalian cells.

In an alternative embodiment, a codon optimized polynucleotide which encodes a Therapeutic protein portion of an albumin fusion protein does not hybridize to the wild type polynucleotide encoding the Therapeutic protein under stringent hybridization conditions as described herein. In a further embodiment, a codon optimized polynucleotide which encodes an albumin portion of an albumin fusion protein does not hybridize to the wild type polynucleotide encoding the albumin protein under stringent hybridization conditions as described herein. In another embodiment, a codon optimized polynucleotide which encodes an albumin fusion protein does not hybridize to the wild type polynucleotide encoding the Therapeutic protein portion or the albumin protein portion under stringent hybridization conditions as described herein.

In an additional embodiment, a polynucleotide which encodes a Therapeutic protein portion of an albumin fusion protein does not comprise, or alternatively consist of, the naturally occurring sequence of that Therapeutic protein. In a further embodiment, a polynucleotide which encodes an albumin protein portion of an albumin fusion protein does not comprise, or alternatively consist of, the naturally occurring sequence of albumin protein. In an alternative embodiment, a polynucleotide which encodes an albumin fusion protein does not comprise, or alternatively consist of, the naturally occurring sequence of a Therapeutic protein portion or the albumin protein portion.

Naturally occurring variants are called “allelic variants,” and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. (Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985)). These allelic variants can vary at either the polynucleotide and/or polypeptide level and are included in the present invention. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis.

Using known methods of protein engineering and recombinant DNA technology, variants may be generated to improve or alter the characteristics of the polypeptides of the present invention. For instance, one or more amino acids can be deleted from the N-terminus or C-terminus of the polypeptide of the present invention without substantial loss of biological function. As an example, Ron et al. (J. Biol. Chem. 268: 2984-2988 (1993)) reported variant KGF proteins having heparin binding activity even after deleting 3, 8, or 27 amino-terminal amino acid residues. Similarly, Interferon gamma exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein. (Dobeli et al., J. Biotechnology 7:199-216 (1988).)

Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle and coworkers (J. Biol. Chem. 268:22105-22111 (1993)) conducted extensive mutational analysis of human cytokine IL-1a. They used random mutagenesis to generate over 3,500 individual IL-1a mutants that averaged 2.5 amino acid changes per variant over the entire length of the molecule. Multiple mutations were examined at every possible amino acid position. The investigators found that “[m]ost of the molecule could be altered with little effect on either [binding or biological activity].” In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity from wild-type.

Furthermore, even if deleting one or more amino acids from the N-terminus or C-terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking N- or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art.

Thus, the invention further includes polypeptide variants which have a functional activity (e.g., biological activity and/or therapeutic activity). In one embodiment, the invention provides variants of albumin fusion proteins that have a functional activity (e.g., biological activity and/or therapeutic activity) that corresponds to one or more biological and/or therapeutic activities of the Therapeutic protein corresponding to the Therapeutic protein portion of the albumin fusion protein. In another embodiment, the invention provides variants of albumin fusion proteins that have a functional activity (e.g., biological activity and/or therapeutic activity) that corresponds to one or more biological and/or therapeutic activities of the Therapeutic protein corresponding to the Therapeutic protein portion of the albumin fusion protein. Such variants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as have little effect on activity. Polynucleotides encoding such variants are also encompassed by the invention.

In preferred embodiments, the variants of the invention have conservative substitutions. By “conservative substitutions” is intended swaps within groups such as replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile; replacement of the hydroxyl residues Ser and Thr; replacement of the acidic residues Asp and Glu; replacement of the amide residues Asn and Gin, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp, and replacement of the small-sized amino acids Ala, Ser, Thr, Met, and Gly.

Guidance concerning how to make phenotypically silent amino acid substitutions is provided, for example, in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that there are two main strategies for studying the tolerance of an amino acid sequence to change.

The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein.

The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (introduction of single alanine mutations at every residue in the molecule) can be used. See Cunningham and Wells, Science 244:1081-1085 (1989). The resulting mutant molecules can then be tested for biological activity.

As the authors state, these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile; replacement of the hydroxyl residues Ser and Thr; replacement of the acidic residues Asp and Glu; replacement of the amide residues Asn and Gln, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp, and replacement of the small-sized amino acids Ala, Ser, Thr, Met, and Gly. Besides conservative amino acid substitution, variants of the present invention include (i) polypeptides containing substitutions of one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) polypeptides containing substitutions of one or more of the amino acid residues having a substituent group, or (iii) polypeptides which have been fused with or chemically conjugated to another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), (iv) polypeptide containing additional amino acids, such as, for example, an IgG Fc fusion region peptide. Such variant polypeptides are deemed to be within the scope of those skilled in the art from the teachings herein.

For example, polypeptide variants containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as less aggregation. Aggregation of pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity. See Pinckard et al., Clin. Exp. Immunol. 2:331-340 (1967); Robbins et al., Diabetes 36: 838-845 (1987); Cleland et al., Crit. Rev. Therapeutic Drug Carrier Systems 10:307-377 (1993).

In specific embodiments, the polypeptides of the invention comprise, or alternatively, consist of, fragments or variants of the amino acid sequence of an albumin fusion protein, the amino acid sequence of a Therapeutic protein and/or human serum albumin, wherein the fragments or variants have 1-5, 5-10.5-25, 5-50, 10-50 or 50-150, amino acid residue additions, substitutions, and/or deletions when compared to the reference amino acid sequence. In preferred embodiments, the amino acid substitutions are conservative. Nucleic acids encoding these polypeptides are also encompassed by the invention.

The polypeptide of the present invention can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The polypeptides may be modified by either natural processes, such as post-translational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, PROTEINS—STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); POST-TRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth. Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992)).

Functional Activity

“A polypeptide having functional activity” refers to a polypeptide capable of displaying one or more known functional activities associated with the full-length, pro-protein, and/or mature form of a Therapeutic protein. Such functional activities include, but are not limited to, biological activity, antigenicity [ability to bind (or compete with a polypeptide for binding) to an anti-polypeptide antibody], immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide.

“A polypeptide having biological activity” refers to a polypeptide exhibiting activity similar to, but not necessarily identical to, an activity of a Therapeutic protein of the present invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the polypeptide of the present invention (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the polypeptide of the present invention).

In preferred embodiments, an albumin fusion protein of the invention has at least one biological and/or therapeutic activity associated with the Therapeutic protein portion (or fragment or variant thereof) when it is not fused to albumin.

The albumin fusion proteins of the invention can be assayed for functional activity (e.g., biological activity) using or routinely modifying assays known in the art, as well as assays described herein. Additionally, one of skill in the art may routinely assay fragments of a Therapeutic protein corresponding to a Therapeutic protein portion of an albumin fusion protein, for activity using assays referenced in its corresponding row of Table 1 (e.g., in column 3 of Table 1). Further, one of skill in the art may routinely assay fragments of an albumin protein corresponding to an albumin protein portion of an albumin fusion protein, for activity using assays known in the art and/or as described in the Examples section below.

For example, in one embodiment where one is assaying for the ability of an albumin fusion protein to bind or compete with a Therapeutic protein for binding to an anti-Therapeutic polypeptide antibody and/or anti-albumin antibody, various immunoassays known in the art can be used, including but not limited to, competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoradiometric assays, gel diffusion precipitation reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

In a preferred embodiment, where a binding partner (e.g., a receptor or a ligand) of a Therapeutic protein is identified, binding to that binding partner by an albumin fusion protein which comprises that Therapeutic protein as the Therapeutic protein portion of the fusion can be assayed, e.g., by means well-known in the art, such as, for example, reducing and non-reducing gel chromatography, protein affinity chromatography, and affinity blotting. See generally, Phizicky et al., Microbiol. Rev. 59:94-123 (1995). In another embodiment, the ability of physiological correlates of an albumin fusion protein to bind to a substrate(s) of the Therapeutic polypeptide corresponding to the Therapeutic protein portion of the fusion can be routinely assayed using techniques known in the art.

In an alternative embodiment, where the ability of an albumin fusion protein to multimerize is being evaluated, association with other components of the multimer can be assayed, e.g., by means well-known in the art, such as, for example, reducing and non-reducing gel chromatography, protein affinity chromatography, and affinity blotting. See generally, Phizicky et al., supra.

In preferred embodiments, an albumin fusion protein comprising all or a portion of an antibody that binds a Therapeutic protein, has at least one biological and/or therapeutic activity (e.g., to specifically bind a polypeptide or epitope) associated with the antibody that binds a Therapeutic protein (or fragment or variant thereof) when it is not fused to albumin. In other preferred embodiments, the biological activity and/or therapeutic activity of an albumin fusion protein comprising all or a portion of an antibody that binds a Therapeutic protein is the inhibition (i.e., antagonism) or activation (i.e., agonism) of one or more of the biological activities and/or therapeutic activities associated with the polypeptide that is specifically bound by antibody that binds a Therapeutic protein.

Albumin fusion proteins comprising at least a fragment or variant of an antibody that binds a Therapeutic protein may be characterized in a variety of ways. In particular, albumin fusion proteins comprising at least a fragment or variant of an antibody that binds a Therapeutic protein may be assayed for the ability to specifically bind to the same antigens specifically bound by the antibody that binds a Therapeutic protein corresponding to the Therapeutic protein portion of the albumin fusion protein using techniques described herein or routinely modifying techniques known in the art.

Assays for the ability of the albumin fusion proteins (e.g., comprising at least a fragment or variant of an antibody that binds a Therapeutic protein) to (specifically) bind a specific protein or epitope may be performed in solution (e.g., Houghten, Bio/Techniques 13:412-421(1992)), on beads (e.g., Lam, Nature 354:82-84 (1991)), on chips (e.g., Fodor, Nature 364:555-556 (1993)), on bacteria (e.g., U.S. Pat. No. 5,223,409), on spores (e.g., U.S. Pat. Nos. 5,571,698; 5,403,484; and 5,223,409), on plasmids (e.g., Cull et al., Proc. Natl. Acad. Sci. USA 89:1865-1869 (1992)) or on phage (e.g., Scott and Smith, Science 249:386-390 (1990); Devlin, Science 249:404-406 (1990); Cwirla et al. Proc. Natl. Acad. Sci. USA 87:6378-6382 (1990); and Felici, J. Mol. Biol. 222:301-310 (1991)) (each of these references is incorporated herein in its entirety by reference). Albumin fusion proteins comprising at least a fragment or variant of a Therapeutic antibody may also be assayed for their specificity and affinity for a specific protein or epitope using or routinely modifying techniques described herein or otherwise known in the art.

The albumin fusion proteins comprising at least a fragment or variant of an antibody that binds a Therapeutic protein may be assayed for cross-reactivity with other antigens (e.g., molecules that have sequence/structure conservation with the molecule(s) specifically bound by the antibody that binds a Therapeutic protein (or fragment or variant thereof) corresponding to the Therapeutic protein portion of the albumin fusion protein of the invention) by any method known in the art.

Immunoassays which can be used to analyze (immunospecific) binding and cross-reactivity include, but are not limited to, competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays, to name but a few. Such assays are routine and well known in the art (see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, which is incorporated by reference herein in its entirety). Exemplary immunoassays are described briefly below (but are not intended by way of limitation).

Immunoprecipitation protocols generally comprise lysing a population of cells in a lysis buffer such as RIPA buffer (1% NP-40 or Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 0.01 M sodium phosphate at pH 7.2, 1% Trasylol) supplemented with protein phosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin, sodium vanadate), adding the albumin fusion protein of the invention (e.g., comprising at least a fragment or variant of an antibody that binds a Therapeutic protein) to the cell lysate, incubating for a period of time (e.g., 1 to 4 hours) at 40 degrees C., adding sepharose beads coupled to an anti-albumin antibody, for example, to the cell lysate, incubating for about an hour or more at 40 degrees C., washing the beads in lysis buffer and resuspending the beads in SDS/sample buffer. The ability of the albumin fusion protein to immunoprecipitate a particular antigen can be assessed by, e.g., western blot analysis. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the binding of the albumin fusion protein to an antigen and decrease the background (e.g., pre-clearing the cell lysate with sepharose beads). For further discussion regarding immunoprecipitation protocols see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 10.16.1.

Western blot analysis generally comprises preparing protein samples, electrophoresis of the protein samples in a polyacrylamide gel (e.g., 8%-20% SDS-PAGE depending on the molecular weight of the antigen), transferring the protein sample from the polyacrylamide gel to a membrane such as nitrocellulose, PVDF or nylon, blocking the membrane in blocking solution (e.g., PBS with 3% BSA or non-fat milk), washing the membrane in washing buffer (e.g., PBS-Tween 20), applying the albumin fusion protein of the invention (diluted in blocking buffer) to the membrane, washing the membrane in washing buffer, applying a secondary antibody (which recognizes the albumin fusion protein, e.g., an anti-human serum albumin antibody) conjugated to an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) or radioactive molecule (e.g., 32P or 125I) diluted in blocking buffer, washing the membrane in wash buffer, and detecting the presence of the antigen. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected and to reduce the background noise. For further discussion regarding western blot protocols see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 10.8.1.

ELISAs comprise preparing antigen, coating the well of a 96-well microtiter plate with the antigen, washing away antigen that did not bind the wells, adding the albumin fusion protein (e.g., comprising at least a fragment or variant of an antibody that binds a Therapeutic protein) of the invention conjugated to a detectable compound such as an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) to the wells and incubating for a period of time, washing away unbound or non-specifically bound albumin fusion proteins, and detecting the presence of the albumin fusion proteins specifically bound to the antigen coating the well. In ELISAs the albumin fusion protein does not have to be conjugated to a detectable compound; instead, a second antibody (which recognizes albumin fusion protein) conjugated to a detectable compound may be added to the well. Further, instead of coating the well with the antigen, the albumin fusion protein may be coated to the well. In this case, the detectable molecule could be the antigen conjugated to a detectable compound such as an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase). One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected as well as other variations of ELISAs known in the art. For further discussion regarding ELISAs see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 11.2.1.

The binding affinity of an albumin fusion protein to a protein, antigen, or epitope and the off-rate of an albumin fusion protein-protein/antigen/epitope interaction can be determined by competitive binding assays. One example of a competitive binding assay is a radioimmunoassay comprising the incubation of labeled antigen (e.g., 3H or 125I) with the albumin fusion protein of the invention in the presence of increasing amounts of unlabeled antigen, and the detection of the antibody bound to the labeled antigen. The affinity of the albumin fusion protein for a specific protein, antigen, or epitope and the binding off-rates can be determined from the data by Scatchard plot analysis. Competition with a second protein that binds the same protein, antigen or epitope as the albumin fusion protein, can also be determined using radioimmunoassays. In this case, the protein, antigen or epitope is incubated with an albumin fusion protein conjugated to a labeled compound (e.g., 3H or 125I) in the presence of increasing amounts of an unlabeled second protein that binds the same protein, antigen, or epitope as the albumin fusion protein of the invention.

In a preferred embodiment, BIAcore kinetic analysis is used to determine the binding on and off rates of albumin fusion proteins of the invention to a protein, antigen or epitope. BIAcore kinetic analysis comprises analyzing the binding and dissociation of albumin fusion proteins, or specific polypeptides, antigens or epitopes from chips with immobilized specific polypeptides, antigens or epitopes or albumin fusion proteins, respectively, on their surface.

Antibodies that bind a Therapeutic protein corresponding to the Therapeutic protein portion of an albumin fusion protein may also be described or specified in terms of their binding affinity for a given protein or antigen, preferably the antigen which they specifically bind. Preferred binding affinities include those with a dissociation constant or Kd less than 5×10−2 M, 10−2 M, 5×10−3 M, 10−3 M, 5×10−4 M, 10−4 M. More preferred binding affinities include those with a dissociation constant or Kd less than 5×10−5 M, 10−5 M, 5×10−6 M, 10−6 M, 5×10−7 M, 10−7 M, 5×10−8 M or 10−8 M. Even more preferred binding affinities include those with a dissociation constant or Kd less than 5×10−9 M, 10−9 M, 5×10−10 M, 10−10 M, 5×10−11 M, 10−11 M, 5×10−12 M, 10−12 M, 5×10−13 M, 10−13 M, 5×10−14 M, 10−14 M, 5×10−15 M, or 10−15 M. In preferred embodiments, albumin fusion proteins comprising at least a fragment or variant of an antibody that binds a Therapeutic protein, has an affinity for a given protein or epitope similar to that of the corresponding antibody (not fused to albumin) that binds a Therapeutic protein, taking into account the valency of the albumin fusion protein (comprising at least a fragment or variant of an antibody that binds a Therapeutic protein) and the valency of the corresponding antibody. In addition, assays described herein (see Examples and Table 1) and otherwise known in the art may routinely be applied to measure the ability of albumin fusion proteins and fragments, variants and derivatives thereof to elicit biological activity and/or Therapeutic activity (either in vitro or in vivo) related to either the Therapeutic protein portion and/or albumin portion of the albumin fusion protein. Other methods will be known to the skilled artisan and are within the scope of the invention.

Albumin

As described above, an albumin fusion protein of the invention comprises at least a fragment or variant of a Therapeutic protein and at least a fragment or variant of human serum albumin. which are associated with one another, preferably by genetic fusion.

An additional embodiment comprises at least a fragment or variant of a Therapeutic protein and at least a fragment or variant of human serum albumin, which are linked to one another by chemical conjugation. The terms, human serum albumin (HSA) and human albumin (HA) are used interchangeably herein. The terms, “albumin and “serum albumin” are broader, and encompass human serum albumin (and fragments and variants thereof) as well as albumin from other species (and fragments and variants thereof).

As used herein, “albumin” refers collectively to albumin protein or amino acid sequence, or an albumin fragment or variant, having one or more functional activities (e.g., biological activities) of albumin. In particular, “albumin” refers to human albumin or fragments thereof (see for example, EP 201 239, EP 322 094 WO 97/24445, WO95/23857) especially the mature form of human albumin as shown in FIG. 1 and SEQ ID NO: 1038, or albumin from other vertebrates or fragments thereof, or analogs or variants of these molecules or fragments thereof.

In preferred embodiments, the human serum albumin protein used in the albumin fusion proteins of the invention contains one or both of the following sets of point mutations with reference to SEQ ID NO: 1038: Leu-407 to Ala, Leu-408 to Val, Val-409 to Ala, and Arg-410 to Ala; or Arg-410 to A, Lys-413 to Gln, and Lys-414 to Gln (see, e.g., International Publication No. WO95/23857, hereby incorporated in its entirety by reference herein). In even more preferred embodiments, albumin fusion proteins of the invention that contain one or both of above-described sets of point mutations have improved stability/resistance to yeast Yap3p proteolytic cleavage, allowing increased production of recombinant albumin fusion proteins expressed in yeast host cells.

As used herein, a portion of albumin sufficient to prolong the therapeutic activity or shelf-life of the Therapeutic protein refers to a portion of albumin sufficient in length or structure to stabilize or prolong the therapeutic activity of the protein so that the shelf life of the Therapeutic protein portion of the albumin fusion protein is prolonged or extended compared to the shelf-life in the non-fusion state. The albumin portion of the albumin fusion proteins may comprise the full length of the HA sequence as described above, or may include one or more fragments thereof that are capable of stabilizing or prolonging the therapeutic activity. Such fragments may be of 10 or more amino acids in length or may include about 15, 20, 25, 30, 50, or more contiguous amino acids from the HA sequence or may include part or all of specific domains of HA. For instance, one or more fragments of HA spanning the first two immunoglobulin-like domains may be used. In a preferred embodiment, the HA fragment is the mature form of HA.

The albumin portion of the albumin fusion proteins of the invention may be a variant of normal HA. The Therapeutic protein portion of the albumin fusion proteins of the invention may also be variants of the Therapeutic proteins as described herein. The term “variants” includes insertions, deletions and substitutions, either conservative or non conservative, where such changes do not substantially alter one or more of the oncotic, useful ligand-binding and non-immunogenic properties of albumin, or the active site, or active domain which confers the therapeutic activities of the Therapeutic proteins.

In particular, the albumin fusion proteins of the invention may include naturally occurring polymorphic variants of human albumin and fragments of human albumin, for example those fragments disclosed in EP 322 094 (namely HA (Pn), where n is 369 to 419). The albumin may be derived from any vertebrate, especially any mammal, for example human, cow, sheep, or pig. Non-mammalian albumins include, but are not limited to, hen and salmon. The albumin portion of the albumin fusion protein may be from a different animal than the Therapeutic protein portion.

Generally speaking, an HA fragment or variant will be at least 100 amino acids long, preferably at least 150 amino acids long. The HA variant may consist of or alternatively comprise at least one whole domain of HA, for example domains 1 (amino acids 1-194 of SEQ ID NO: 1038), domain 2 (amino acids 195-387 of SEQ ID NO: 1038), domain 3 (amino acids 388-585 of SEQ ID NO: 1038), domains 1 and 2 (1-387 of SEQ ID NO: 1038), domains 2 and 3 (195-585 of SEQ ID NO: 1038) or domains 1 and 3 (amino acids I-194 of SEQ ID NO: 1038 and amino acids 388-585 of SEQ ID NO: 1038). Each domain is itself made up of two homologous subdomains namely 1-105, 120-194, 195-291, 316-387, 388-491 and 512-585, with flexible inter-subdomain linker regions comprising residues Lys106 to Glu119, Glu292 to Val315 and Glu492 to Ala511:

Preferably, the albumin portion of an albumin fusion protein of the invention comprises at least one subdomain or domain of HA or conservative modifications thereof. If the fusion is based on subdomains, some or all of the adjacent linker is preferably used to link to the Therapeutic protein moiety.

Antibodies that Specifically Bind Therapeutic Proteins are Also Therapeutic Proteins

The present invention also encompasses albumin fusion proteins that comprise at least a fragment or variant of an antibody that specifically binds a Therapeutic protein disclosed in Table 1. It is specifically contemplated that the term “Therapeutic protein” encompasses antibodies that bind a Therapeutic protein (e.g., as Described in column 1 of Table 1) and fragments and variants thereof. Thus an albumin fusion protein of the invention may contain at least a fragment or variant of a Therapeutic protein, and/or at least a fragment or variant of an antibody that binds a Therapeutic protein.

Antibody Structure and Background

The basic antibody structural unit is known to comprise a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region f about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. See generally, Fundamental Immunology Chapters 3-5 (Paul, W., ed., 4th ed. Raven Press, N.Y. (1998)) (incorporated by reference in its entirety for all purposes). The variable regions of each light/heavy chain pair form the antibody binding site.

Thus, an intact IgG antibody has two binding sites. Except in bifunctional or bispecific antibodies, the two binding sites are the same.

The chains all exhibit the same general structure of relatively conserved framework regions (FR) joined by three hypervariable regions, also called complementarity determining regions or CDRs. The CDR regions, in general, are the portions of the antibody which make contact with the antigen and determine its specificity. The CDRs from the heavy and the light chains of each pair are aligned by the framework regions, enabling binding to a specific epitope. From N-terminal to C-terminal, both light and heavy chains variable regions comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The variable regions are connected to the heavy or light chain constant region. The assignment of amino acids to each domain is in accordance with the definitions of Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk J Mol. Biol. 196:901-917 (1987); Chothia et al. Nature 342:878-883 (1989).

As used herein, “antibody” refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that specifically binds an antigen (e.g., a molecule containing one or more CDR regions of an antibody). Antibodies that may correspond to a Therapeutic protein portion of an albumin fusion protein include, but are not limited to, monoclonal, multispecific, human, humanized or chimeric antibodies, single chain antibodies (e.g., single chain Fvs), Fab fragments, F(ab′) fragments, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies specific to antibodies of the invention), and epitope-binding fragments of any of the above (e.g., VH domains, VL domains, or one or more CDR regions).

Antibodies that Bind Therapeutic Proteins

The present invention encompasses albumin fusion proteins that comprise at least a fragment or variant of an antibody that binds a Therapeutic Protein (e.g., as disclosed in Table 1) or fragment or variant thereof.

Antibodies that bind a Therapeutic protein (or fragment or variant thereof) may be from any animal origin, including birds and mammals. Preferably, the antibodies are human, murine (e.g., mouse and rat), donkey, sheep, rabbit, goat, guinea pig, camel, horse, or chicken antibodies. Most preferably, the antibodies are human antibodies. As used herein, “human” antibodies include antibodies having the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries and xenomice or other organisms that have been genetically engineered to produce human antibodies.

The antibody molecules that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule. In preferred embodiments, the antibody molecules that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein are IgG1. In other preferred embodiments, the immunoglobulin molecules that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein are IgG2. In other preferred embodiments, the immunoglobulin molecules that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein are IgG4.

Most preferably the antibodies that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein are human antigen-binding antibody fragments of the present invention and include, but are not limited to, Fab, Fab′ and F(ab′)2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a VL or VH domain. Antigen-binding antibody fragments, including single-chain antibodies, may comprise the variable region(s) alone or in combination with the entirety or a portion of the following: hinge region, CH1, CH2, and CH3 domains.

The antibodies that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein may be monospecific, bispecific, trispecific or of greater multispecificity. Multispecific antibodies may be specific for different epitopes of a Therapeutic protein or may be specific for both a Therapeutic protein as well as for a heterologous epitope, such as a heterologous polypeptide or solid support material. See, e.g., PCT publications WO 93/17715; WO 92/08802; WO 91/00360; WO 92/05793; Tutt, et al., J. Immunol. 147:60-69 (1991); U.S. Pat. Nos. 4,474,893; 4,714,681; 4,925,648; 5,573,920; 5,601,819; Kostelny et al., J. Immunol. 148:1547-1553 (1992).

Antibodies that bind a Therapeutic protein (or fragment or variant thereof) may be bispecific or bifunctional which means that the antibody is an artificial hybrid antibody having two different heavy/light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab′ fragments. See, e.g., Songsivilai & Lachmann Clin. Exp. Immunol. 79: 315-321 (1990), Kostelny et al. J Immunol. 148:1547 1553 (1992). In addition, bispecific antibodies may be formed as “diabodies” (Holliger et al. “‘Diabodies’: small bivalent and bispecific antibody fragments” PNAS USA 90:6444-6448 (1993)) or “Janusins” (Traunecker et al. “Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells” EMBO J 10:3655-3659 (1991) and Traunecker et al. “Janusin: new molecular design for bispecific reagents” Int J Cancer Suppl 7:51-52 (1992)).

The present invention also provides albumin fusion proteins that comprise, fragments or variants (including derivatives) of an antibody described herein or known elsewhere in the art. Standard techniques known to those of skill in the art can be used to introduce mutations in the nucleotide sequence encoding a molecule of the invention, including, for example, site-directed mutagenesis and PCR-mediated mutagenesis which result in amino acid substitutions. Preferably, the variants (including derivatives) encode less than 50 amino acid substitutions, less than 40 amino acid substitutions, less than 30 amino acid substitutions, less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the reference VH domain, VHCDR1, VHCDR2, VHCDR3, VL domain, VLCDR1, VLCDR2, or VLCDR3. In specific embodiments, the variants encode substitutions of VHCDR3. In a preferred embodiment, the variants have conservative amino acid substitutions at one or more predicted non-essential amino acid residues.

Antibodies that bind to a Therapeutic protein and that may correspond to a Therapeutic protein portion of an albumin fusion protein may be described or specified