Claims
- 1. A method for preparing a mixture of intact and single-clipped isoforms of biologically active recombinant PDGF-BB (rPDGF-BB) dimeric protein from a mixture of isoforms of said protein obtained from a cell culture, wherein the rPDGF-B polypeptides in said rPDGF-BB dimeric protein are native-sequence or variant rPDGF-B polypeptides that exhibit at least about 80% sequence identity to native-sequence rPDGF-B polypeptides, said method comprising loading said mixture of isoforms onto a TSK sulfopropyl cation exchange chromatography column, washing said mixture of isoforms from said column with a linear gradient buffer, discarding an eluted fraction comprising a double-clipped isoform, and collecting an eluted fraction comprising intact and single-clipped isoforms.
- 2. The method of claim 1, wherein the rPDGF-B polypeptides are native-sequence polypeptides.
- 3. The method of claim 2, wherein the rPDGF-B polypeptides have the amino acid sequences of human rPDGF-B polypeptides.
- 4. The method of claim 1, wherein said rPDGF-BB is produced in a yeast host cell.
- 5. The method of claim 4, wherein said yeast host cell is a Saccharomyces yeast host cell.
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application is a Divisional of U.S. application Ser. No. 08/989,250, filed Dec. 12, 1997 (now U.S. Pat. No. 6,083,910), which claims the benefit of U.S. Provisional Application Serial No. 60/032,720, filed Dec. 13, 1996, the contents of which are herein incorporated by reference in their entirety.
US Referenced Citations (3)
Foreign Referenced Citations (1)
Number |
Date |
Country |
0 259 632 |
Aug 1987 |
EP |
Non-Patent Literature Citations (2)
Entry |
Cook et al., “Purification and Analysis of Proteinase-Resistant Mutants of Recombinant Platelet-Derived Growth Factor-BB Exhibiting Improved Biological Activity,” The Biochemical Jouranl, (Jan. 1992) vol. 281, 1, pp. 56-65. |
Oefner et al., “Crystal Structure of Human Platelet-Derived Growth Factor BB,” EMBO Journal (1992), vol. 11, pp. 3921-3926. |
Provisional Applications (1)
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Number |
Date |
Country |
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60/032720 |
Dec 1996 |
US |