Isolated polynucleotide that encodes an ActRIIa-Fc fusion polypeptide
In certain aspects, the present invention provides compositions and methods for promoting bone growth and increasing bone density.
1. An isolated polynucleotide comprising a first nucleic acid sequence that hybridizes to a second nucleic acid sequence that is complementary to the sequence of SEQ ID NO:14 in the presence of 6.0× sodium chloride/sodium citrate (SSC) at 45° C. followed by a wash of 0.2×SSC at 65° C., wherein the first nucleic acid sequence encodes an ActRIIa-Fc fusion polypeptide that has the N-terminal sequence ILGRSETQE (SEQ ID NO:11).
2. The isolated polynucleotide of claim 1 , wherein the nucleic acid sequence comprises a coding sequence for a polypeptide comprising the amino acid sequence of SEQ ID NO:7.
3. The isolated polynucleotide of claim 1 , wherein the polynucleotide comprises the nucleic acid sequence of SEQ ID NO:14.
4. A recombinant polynucleotide comprising a promoter sequence operably linked to the polynucleotide of claim 1 .
5. A cell comprising the recombinant polynucleotide of claim 4 .
6. The cell of claim 5 , wherein the cell is a mammalian cell.
7. The cell of claim 6 , wherein the cell is a Chinese Hamster Ovary (CHO) cell.
8. A vector comprising at least one regulatory sequence operably linked to the polynucleotide of claim 1 .
9. A cell comprising the vector of claim 8 .
10. The cell of claim 9 , wherein the cell is a mammalian cell.
11. The cell of claim 10 , wherein the cell is a CHO cell.
12. A method of making an ActRIIa-Fc fusion polypeptide, wherein the method comprises:
i) providing a cell comprising the polynucleotide of claim 1 ;
ii) culturing the cell under conditions suitable for expression of the ActRIIa-Fc fusion polypeptide encoded by the recombinant polynucleotide; and
iii) recovering the expressed ActRIIa-Fc fusion polypeptide.
13. The method of claim 12 , wherein the cell is a CHO cell.
14. The method of claim 12 , wherein the polynucleotide sequence comprises a coding sequence for a polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:7.
15. The method of claim 12 , wherein the polynucleotide sequence comprises a coding sequence for a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:7.
16. The method of claim 12 , wherein the polynucleotide sequence comprises a coding sequence for a polypeptide comprising the amino acid sequence of SEQ ID NO:7.
17. The method of claim 12 , wherein the polynucleotide comprises the nucleic acid sequence of SEQ ID NO:14.
18. The method of claim 12 , wherein the encoded polypeptide binds to activin and/or GDF11.
19. The method of claim 18 , wherein the encoded polypeptide binds to activin A.
20. The method of claim 18 , wherein the encoded polypeptide binds to activin B.
21. The method of claim 18 , wherein the encoded polypeptide binds to GDF11.
22. The method of claim 18 , wherein the encoded polypeptide binds to activin and GDF11.
23. The method of claim 22 , wherein the encoded polypeptide binds to activin A.
24. The method of claim 22 , wherein the encoded polypeptide binds to activin B.
25. The isolated polynucleotide of claim 1 , wherein the polynucleotide sequence comprises a coding sequence for a polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:7.
26. The isolated polynucleotide of claim 1 , wherein the polynucleotide sequence comprises a coding sequence for a polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:7.
27. The isolated polynucleotide of claim 1 , wherein the encoded polypeptide binds to activin and/or GDF11.
28. The isolated polynucleotide of claim 27 , wherein the encoded polypeptide binds to activin A.
29. The isolated polynucleotide of claim 27 , wherein the encoded polypeptide binds to activin B.
30. The isolated polynucleotide of claim 27 , wherein the encoded polypeptide binds to GDF11.
31. The isolated polynucleotide of claim 27 , wherein the encoded polypeptide binds to activin and GDF11.
32. The isolated polynucleotide of claim 31 , wherein the encoded polypeptide binds to activin A.
33. The isolated polynucleotide of claim 31 , wherein the encoded polypeptide binds to activin B.