Claims
- 1. An isolated polypeptide comprising:
a first C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 31-35, 52-54, 60-63, 67-69, 73-84, 89-95, 101-108, 112-126, 131-136 and 150-154 of SEQ ID NO:2, and a cysteine residue corresponding to residue 70 of SEQ ID NO:2; and a second C1q domain joined to the carboxy terminal of said first C1q domain, said second C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 179-183, 206-208, 214-217, 221-223, 227-238, 243-249, 254-262, 267-278, 283-288 and 305-309 of SEQ ID NO:2, and a cysteine residue corresponding to residue 223, a glycine residue corresponding to residue 228 of SEQ ID NO:2.
- 2. An isolated polypeptide according to claim 1, further comprising a secretory signal sequence.
- 3. An isolated polypeptide according to claim 2, where said secretory signal sequence comprises amino acid residues 1-16 of SEQ ID NO:2
- 4. An isolated polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 17 to 329 of SEQ ID NO:2, wherein said sequence comprises:
a first C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 31-35, 52-54, 60-63, 67-69, 73-84, 89-95, 101-108, 112-126, 131-136 and 150-154 of SEQ ID NO:2, and a cysteine residue corresponding to residue 70 of SEQ ID NO:2; and a second C1q domain joined to the carboxy terminal of said first C1q domain, said second C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 179-183, 206-208, 214-217, 221-223, 227-238, 243-249, 254-262, 267-278, 283-288 and 305-309 of SEQ ID NO:2, and a cysteine residue corresponding to residue 223, a glycine residue corresponding to residue 228 of SEQ ID NO:2.
- 5. An isolated polypeptide according to claim 4, wherein said polypeptide is at least 90% identical in amino acid sequence to residues 17 to 329 of SEQ ID NO:2.
- 6. An isolated polypeptide of claim 5, wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=blosum62, with other parameters set as default.
- 7. An isolated polypeptide according to claim 5, wherein any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions.
- 8. An isolated polypeptide according to claim 5, wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.
- 9. An isolated polypeptide according to claim 5, wherein said first C1q domain comprises amino acid residues 17-159 of SEQ ID NO:2.
- 10. An isolated polypeptide according to claim 5, wherein said second C1q domain comprises amino acid residues 160-328 of SEQ ID NO:2.
- 11. An isolated polypeptide according to claim 5, wherein said polypeptide comprises residues 17-328 of SEQ ID NO:2.
- 12. An isolated polypeptide according to claim 4, covalently linked at the amino or carboxyl terminus to a moiety selected from the group consisting of affinity tags, toxins, radionucleotides, enzymes and fluorophores.
- 13. An isolated polypeptide comprising:
a signal sequence; a first C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 31-35, 52-54, 60-63, 67-69, 73-84, 89-95, 101-108, 112-126, 131-136 and 150-154 of SEQ ID NO:2 and a cysteine residue corresponding to amino acid residue 70 of SEQ ID NO:2; and a second C1q domain joined to the carboxy terminal of said first C1q domain, said second C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 179-183, 206-208, 214-217, 221-223, 227-238, 243-249, 254-262, 267-278, 283-288 and 305-309 of SEQ ID NO:2 and a cysteine residue corresponding to residue 223 of SEQ ID NO:2; wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.
- 14. An isolated polypeptide selected from the group consisting of:
a) a polypeptide consisting of amino acid residue 17 to amino acid residue 159 of SEQ ID NO:2 or b) a polypeptide consisting of amino acid residue 160 to amino acid residue 329 of SEQ ID NO:2.
- 15. A fusion protein consisting essentially of a first portion and a second portion joined by a peptide bond, said first portion consisting of a polypeptide selected from the group consisting of:
a) polypeptide according the claim 1;b) a polypeptide comprising the amino acid sequence of residues 17-159 of SEQ ID NO:2; c) a polypeptide comprising the amino acid sequence of residues 160-329 of SEQ ID NO:2; d) a polypeptide comprising the amino acid sequence of residues 17-329 of SEQ ID NO:2; and said second portion comprising another polypeptide.
- 16. A polypeptide according to claim 1; in combination with a pharmaceutically acceptable vehicle.
- 17. A method of producing an antibody to a polypeptide comprising:
inoculating an animal with a polypeptide selected from the group consisting of:
a) polypeptide according to claim 1;b) a polypeptide comprising the amino acid sequence of residues 17-159 of SEQ ID NO:2; c) a polypeptide comprising the amino acid sequence of residues 160-329 of SEQ ID NO:2; d) a polypeptide comprising the amino acid sequence of residues 17-329 of SEQ ID NO:2; and e) a polypeptide or polypeptide fragment of SEQ ID NO:2; and wherein said polypeptide elicits an immune response in the animal to produce the antibody; and isolating the antibody from the animal.
- 18. An antibody or antibody fragment that specifically binds to a polypeptide according to claim 1.
- 19. An antibody according to claim 18, wherein said antibody is selected from the group consisting of:
a) polyclonal antibody; b) murine monoclonal antibody; c) humanized antibody derived from b); and d) human monoclonal antibody.
- 20. An antibody fragment according to claim 18, wherein said antibody fragment is selected from the group consisting of F(ab′), F(ab), Fab′, Fab, Fv, scFv, and minimal recognition unit.
- 21. An anti-idiotype antibody that specifically binds to said antibody of claim 18.
- 22. An isolated polynucleotide molecule encoding a polypeptide according to claim 1.
- 23. An isolated polynucleotide molecule according to claim 22, wherein said polypeptide further comprises a secretory signal sequence.
- 24. An isolated polynucleotide molecule according to claim 23, where said secretory signal sequence comprises amino acid residues 1-16 of SEQ ID NO:2
- 25. An isolated polynucleotide molecule encoding a polypeptide according to claim 4.
- 26. An isolated polynucleotide molecule according to claim 25, wherein said polypeptide is at least 90% identical in amino acid sequence to residues 17-329 of SEQ ID NO:2.
- 27. An isolated polynucleotide molecule of claim 26, wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=blosum62, with other parameters set as default.
- 28. An isolated polynucleotide molecule according to claim 26, wherein any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions.
- 29. An isolated polynucleotide molecule according to claim 26, wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.
- 30. An isolated polynucleotide molecule according to claim 26, wherein said polynucleotide molecule remains hybridized following stringent wash conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, or the complement of SEQ ID NO:1.
- 31. An isolated polynucleotide molecule according to claim 26, wherein said first C1q domain comprises amino acid residues 17-159 of SEQ ID NO:2.
- 32. An isolated polynucleotide molecule according to claim 26, wherein said second C1q domain comprises amino acid residues 160-329 of SEQ ID NO:2.
- 33. An isolated polynucleotide molecule according to claim 26, wherein said polypeptide comprises residues 17-329 of SEQ ID NO:2.
- 34. An isolated polynucleotide molecule encoding a polypeptide comprising:
a signal sequence;
a first carboxyl-terminal C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 31-35, 52-54, 60-63, 67-69, 73-84, 89-95, 101-108, 112-126, 131-136 and 150-154 of SEQ ID NO:2 and a cysteine residue corresponding to amino acid residue 70 of SEQ ID NO:2; and
a second carboxyl-terminal C1q domain comprising a sequence of SEQ ID NO:3, 10 beta strands corresponding to amino acid residues 179-183, 206-208, 214-217, 221-223, 227-238, 243-249, 254-262, 267-278, 283-288 and 305-309 of SEQ ID NO:2 and a cysteine residue corresponding to residue 223 of SEQ ID NO:2; wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2, and said polynucleotide molecule remains hybridized following stringent wash conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, or the complement of SEQ ID NO:1.
- 35. An isolated polynucleotide molecule selected from the group consisting of,
a) a contiguous sequence of nucleotides from nucleotide 1 to nucleotide 1357 of SEQ ID NO:1; b) a contiguous sequence of nucleotides from nucleotide 210 to nucleotide 1196 of SEQ ID NO:1; c) a contiguous sequence of nucleotides from nucleotide 258 to nucleotide 1196 of SEQ ID NO:1; d) a contiguous sequence of nucleotides from nucleotide 258 to nucleotide 686 of SEQ ID NO:1; e) a polynucleotide encoding a polypeptide consisting of the sequence of amino acid residues 17 to 159 of SEQ ID NO:2; f) a polynucleotide encoding a polypeptide consisting of the sequence of amino acid residues 160 to 329 of SEQ ID NO:2; g) a polynucleotide encoding a polypeptide consisting of the sequence of amino acid residues 17 to 329 of SEQ ID NO:2; h) a polynucleotide that remains hybridized following stringent wash conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:1, or the complement of SEQ ID NO: 1; i) the nucleotide sequences complementary to a), b), c), d), e), f), g) or h) and m) degenerate nucleotide sequences of e), f) or g).
- 36. An isolated polynucleotide molecule encoding a fusion protein consisting essentially of a first portion and a second portion joined by a peptide bond, said first portion is selected from the group consisting of:
a) a polypeptide according to claim 1;b) a polypeptide comprising the amino acid sequence of residues 17-159 of SEQ ID NO:2; c) a polypeptide comprising the amino acid sequence of residues 160-329 of SEQ ID NO:2; and d) a polypeptide comprising the amino acid sequence of residues 17-329 of SEQ ID NO:2; and said second portion comprising another polypeptide.
- 37. An isolated polynucleotide molecule consisting of the sequence of nucleotide 1 to nucleotide 987 of SEQ ID NO:4.
- 38. An expression vector comprising the following operably linked elements:
a transcription promoter; a DNA segment encoding a polypeptide according to claim 1; and a transcription terminator.
- 39. An expression vector according to claim 38, wherein said DNA segment encodes a polypeptide covalently linked at the amino or carboxyl terminus to an affinity tag.
- 40. An expression vector according to claim 38, wherein said DNA segment further encodes a secretory signal sequence operably linked to said polypeptide.
- 41. An expression vector according the claim 40, wherein said secretory signal sequence comprises residues 1-16 of SEQ ID NO:2.
- 42. A cultured cell into which has been introduced an expression vector according to claim 38, wherein said cell expresses said polypeptide encoded by said DNA segment.
- 43. A method of producing a protein comprising:
culturing a cell into which has been introduced an expression vector according to claim 38;whereby said cell expresses said polypeptide encoded by said DNA segment; and recovering said expressed protein.
REFERENCE TO RELATED APPLICATIONS
[0001] This application is related to Provisional Application No. 60/141,928, filed on Jul. 1, 1999. Under 35 U.S.C. §119(e)(1), this application claims benefit of said Provisional Application.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60141928 |
Jul 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09606031 |
Jun 2000 |
US |
Child |
10085167 |
Feb 2002 |
US |