Albumin fusion proteins
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.
1. An albumin fusion protein comprising a Factor IX polypeptide fused to an albumin polypeptide, wherein said albumin polypeptide has the ability to prolong the serum half-life of the fused Factor IX polypeptide compared to the unfused Factor IX polypeptide, wherein said fused Factor IX polypeptide has Factor IX clotting activity, wherein said Factor IX polypeptide is fused at the N-terminus of the albumin polypeptide, wherein said albumin polypeptide is:
(i) a full-length albumin;
(ii) a mature albumin;
(iii) a fragment of albumin; or
(iv) a polypeptide consisting of an amino acid sequence at least 95% identical to SEQ ID NO:18;
and wherein said Factor IX polypeptide is:
a) a full-length Factor IX;
b) an activated Factor IX (Factor IXa);
c) a fragment of Factor IX having a deletion that consists of 1-50 amino acid residues as compared to the sequence of amino acids 1 (Tyr) to 416 (Thr) of SEQ ID NO:46;
d) a polypeptide consisting of an amino acid sequence at least 90% identical to amino acids 1 (Tyr) to 416 (Thr) of SEQ ID NO: 46; or
e) a mature Factor X.
2. The albumin fusion protein of claim 1 , wherein said Factor IX polypeptide is: a), b), d), or e).
3. The albumin fusion protein of claim 1 , wherein said fragment of Factor IX is an N-terminal deletion mutant, a C-terminal deletion mutant, or an N-terminal and C-terminal deletion mutant.
4. The albumin fusion protein of claim 1 , wherein said Factor IX polypeptide is separated from said albumin polypeptide by a peptide linker.
5. The albumin fusion protein of claim 1 , further comprising a secretion leader sequence.
6. The albumin fusion protein of claim 1 , which is non-glycosylated.
7. The albumin fusion protein of claim 1 , which is expressed in yeast.
8. The albumin fusion protein of claim 7 , wherein said yeast is a Saccharomyces cerevisiae.
9. The albumin fusion protein of claim 7 , wherein said yeast is glycosylation deficient.
10. The albumin fusion protein of claim 7 , wherein said yeast is glycosylation and protease deficient.
11. The albumin fusion protein of claim 7 , wherein said fusion protein is encoded by a polynucleotide that is codon-optimized for expression in yeast.
12. The albumin fusion protein of claim 1 , wherein said fusion protein is expressed by a mammalian cell.
13. The albumin fusion protein of claim 12 , wherein said mammalian cell is a COS, CHO (Chinese hamster ovary), or NSO cell.
14. A composition comprising the albumin fusion protein of claim 1 and a pharmaceutically acceptable carrier.
15. The albumin fusion protein of claim 1 , wherein said fragment of albumin comprises amino acids 1 to 387 of SEQ ID NO:18, or amino acids 1 to 194 of SEQ ID NO:18, or amino acids 195 to 387 of SEQ ID NO:18, or amino acids 388 to 585 of SEQ ID NO:18, or amino acids 195 to 585 of SEQ ID NO:18, or amino acids 1 to 194 and amino acids 388 to 585 of SEQ ID NO:18.
16. The albumin fusion protein of claim 1 , wherein said Factor IX polypeptide is selected from:
a) a full-length human Factor IX;
b) an activated human factor IX (Factor IXa);
c) a fragment of a human Factor IX having a deletion that consists of 1-50 amino acid residues as compared to mature human Factor IX;
d) a polypeptide consisting of an amino acid sequence at least 90% identical to amino acids 47-462 of SEQ ID NO: 46; and
e) a mature human Factor IX.
17. The albumin fusion protein of claim 1 , wherein said fragment of Factor IX has a deletion that consists of 1-25 amino acid residues as compared to the sequence of amino acids 1 (Tyr) to 416 (Thr) of SEQ ID NO:46.
18. The albumin fusion protein of claim 1 , wherein said fragment of Factor IX has a deletion that consists of 1-10 amino acid residues as compared to the sequence of amino acids 1 (Tyr) to 416 (Thr) of SEQ ID NO:46.
19. The albumin fusion protein of claim 16 , wherein said fragment of a human Factor IX has a deletion that consists of 1-25 amino acid residues compared to mature human Factor IX.
20. The albumin fusion protein of claim 16 , wherein said fragment of a human Factor IX has a deletion that consists of 1-10 amino acid residues compared to mature human Factor IX.