Methods for producing BMP-7 proteins

• US 5,366,875 A
• Filed: 09/24/1991
• Issued: 11/22/1994
• Est. Priority Date: 07/01/1986
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Wozney, John M., Wang, Elizabeth A., Rosen, Vicki A.
• Primary Examiner(s): Ellis, Joan
• Application Number:
  US07/764,731
• Time in Patent Office: 1,155 Days
• Field of Search: 435/69.1, 435/91, 435/173.2, 435/235.1, 435/320.1, 435/240.1, 536/27, 530/350, 535/9, 535/32, 535/34, 535/57, 535/62, 535/70
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 24/043 Mixtures of macromolecular materials
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C08L 89/00 Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00
• CPC Class Codes: C12N 15/85 for animal cells

Abstract

Purified BMP-7 proteins and processes for producing them are disclosed. The proteins may be used in the treatment of bone and/or cartilage defects and in wound healing and related tissue repair.

Claims

1. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence which encodes the amino acid sequence of SEQ ID NO:

8, culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.

4. The method of claims 1, 2, or 3 wherein said mammalian cells are CHO cells.

2. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence comprising the DNA sequence of SEQ ID NO:

7 which encodes the amino acid sequence of SEQ ID NO;

8, culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.

3. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence comprising the DNA sequence of SEQ ID NO:

7 and sequences which

a) hybridize to the sequence of SEQ ID NO;

7 under stringent hybridization conditions; and

b) express a protein characterized by the ability to induce the formation of cartilage and/or bone formation in the Rosen-modified Sampath Reddi assay

culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.

5. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence which encodes the amino acid sequence of SEQ ID NO:

10, culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.

8. The method of claims 5, 6, or 7 wherein said mammalian cells are CHO cells.

6. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence comprising the DNA sequence of SEQ ID NO:

9 which encodes the amino acid sequence of SEQ ID NO;

10, culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.

7. A method for producing a BMP-7 protein which comprises transforming mammalian cells with an expression vector, said vector having inserted therein a DNA sequence comprising the DNA sequence of SEQ ID NO:

9 and sequences which

a) hybridize to the sequence of SEQ ID NO;

9 under stringent hybridization conditions; and

b) express a protein characterized by the ability to induce the formation of catilage and/or bone in the Rosen-modified Sampath Reddi assay

culturing said cells in a suitable culture medium, and isolating said BMP-7 protein produced by said cells.