Granted Patent B2

US 9,932,379 · App. 15/221,341 · Granted Apr 3, 2018

Inventors: Jasbir Seehra (Lexington, MA); Robert Scott Pearsall (Woburn, MA); Ravindra Kumar (Acton, MA)

Assignee: ACCELERON PHARMA INC.

C07K14/495A61K31/00A61K31/454A61K31/7068A61K38/1709A61K38/179A61K38/1796A61K38/1816A61K38/193A61K39/39533C07K14/475C07K14/505

Abstract

Claims (15)

1. A method for producing a variant ActRIIB polypeptide, comprising:

a) culturing a cell under conditions suitable for expression of the polypeptide, and

b) recovering the polypeptide expressed by the cell;

wherein the cell comprises a nucleic acid; wherein the nucleic acid comprises a nucleic acid sequence that hybridizes to the complement of nucleotides 76-396 of SEQ ID NO: 30 in the presence of 6.0× sodium chloride/sodium citrate (SSC) at 45° C. followed by a wash in 0.2× SSC at 65° C., wherein the nucleic acid sequence encodes a polypeptide that comprises an acidic amino acid at the position corresponding to position 79 of SEQ ID NO: 1.

2. The method of claim 1 , wherein the nucleic acid hybridizes to the complement of the nucleotide sequence of SEQ ID NO: 30 in the presence of 6.0× sodium chloride/sodium citrate (SSC) at 45° C. followed by a wash in 0.2× SSC at 65° C.

3. The method of claim 1 , wherein the nucleic acid comprises nucleotides 76-396 of SEQ ID NO: 30.

4. The method of claim 1 , wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 30.

5. The method of claim 1 , wherein the nucleic acid comprises a nucleotide sequence encoding a polypeptide comprising amino acids 26-132 of SEQ ID NO: 26.

6. The method of claim 1 , wherein the nucleic acid comprises a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 26.

7. The method of claim 1 , wherein the nucleic acid further comprises a nucleotide sequence encoding a tissue plasminogen activator (TPA) leader sequence.

8. The method of claim 1 , wherein the nucleic acid is operably linked to one or more regulatory sequences.

9. The method of claim 1 , wherein the variant polypeptide comprises the amino acid sequence of SEQ ID NO: 28.

10. The method of claim 1 , wherein the cell is a mammalian cell.

11. The method of claim 1 , wherein the cell is a CHO cell.

12. The method of claim 1 , wherein the variant polypeptide comprises an amino acid sequence consisting of SEQ ID NO: 28.

Assignments (1)

Continuity (8)