Claims
- 1. a method for preparing a composition enriched in at least one desired isoform of biologically active recombinant PDGF (rPDGF) dimeric protein or variant thereof from a mixture of isoforms of said rPDGF dimeric protein or variant thereof, said method comprising taking said mixture and enriching said mixture in said desired isoform by reducing amounts of undesired isoforms in said mixture to obtain said composition.
- 2. The method of claim 1, wherein said enriching is accomplished using ion exchange chromatography, reverse phase high-pressure liquid chromatography, or reverse charge capillary zone electrophoresis.
- 3. The method of claim 2, wherein said rPDGF dimeric protein is rPDGF-BB, wherein the rPDGF-B polypeptides in said rPDGF-BB are native-sequence or variant rPDGF-B polypeptides that exhibit at least about 80% sequence identity to native-sequence rPDGF-B polypeptides.
- 4. The method of claim 3, wherein the rPDGF-B polypeptides are native-sequence polypeptides.
- 5. The method of claim 4, wherein the rPDGF-B polypeptides have the amino acid sequences of human rPDGF-B polypeptides.
- 6. The method of claim 3, wherein said rPDGF-BB is produced in a yeast host cell.
- 7. The method of claim 6, wherein said yeast host cell is a Saccharomyces yeast host cell.
- 8. A method for preparing a composition enriched in intact and single-clipped isoforms of biologically active recombinant PDGF (rPDGF)-BB dimeric protein from a mixture of isoforms of said rPDGF-BB dimeric protein, 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 taking said mixture and enriching said mixture in said intact and single-clipped isoforms by reducing amounts of undesired isoforms in said mixture to obtain said composition.
- 9. The method of claim 8, wherein said enriching is accomplished using ion exchange chromatography, reverse phase high-pressure liquid chromatography, or reverse charge capillary zone electrophoresis.
- 10. The method of claim 9, wherein the rPDGF-B polypeptides are native-sequence polypeptides.
- 11. The method of claim 10, wherein the rPDGF-B polypeptides have the amino acid sequences of human rPDGF-B polypeptides.
- 12. The method of claim 9, wherein said rPDGF-BB is produced in a yeast host cell.
- 13. The method of claim 12, wherein said yeast host cell is a Saccharomyces yeast host cell.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation of U.S. application Ser. No. 09/549,290, filed Apr. 14, 2000, which 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. Application Serial No. 60/032,720, filed Dec. 13, 1996, the contents of which are herein incorporated by reference in their entirety.
Provisional Applications (1)
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Number |
Date |
Country |
|
60032720 |
Dec 1996 |
US |
Divisions (1)
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Number |
Date |
Country |
Parent |
08989250 |
Dec 1997 |
US |
Child |
09549290 |
Apr 2000 |
US |
Continuations (1)
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Number |
Date |
Country |
Parent |
09549290 |
Apr 2000 |
US |
Child |
10205693 |
Jul 2002 |
US |