Claims
- 1. A method for purifying factor XIII from a biological fluid, comprising the steps of:
- a) fractionating a biological fluid by anion exchange chromatography to produce a fraction enriched for factor XIII;
- b) adding Na-acetate to the enriched fraction to form a crystalline precipitate;
- c) dissolving the precipitate to form a solution;
- d) fractionating the solution by hydrophobic interaction chromatography to produce a second fraction enriched for factor XIII;
- e) fractionating the second enriched fraction by anion exchange chromatography to produce a third fraction enriched for factor XIII;
- f) adjusting the pH of the third enriched fraction to pH 5.2-6.5 to produce a factor XIII-containing precipitate;
- g) recovering said precipitate;
- h) dissolving the precipitate to form a solution; and
- i) fractionating the solution by gel filtration and collecting a factor XIII-containing peak fraction.
- 2. The method of claim 1 wherein the step of adjusting the pH comprises adding succinic acid to said third enriched fraction.
- 3. The method of claim 1 wherein said biological fluid is a yeast cell lysate.
- 4. The method of claim 1 wherein said factor XIII is human factor XIII.
- 5. The method of claim 1 wherein said factor XIII is recombinant human factor XIII.
- 6. The method of claim 1 wherein said factor XIII is a dimer of a subunits.
- 7. The method of claim 1 wherein the pH of the third enriched fraction is adjusted to 5.8 to produce a factor XIII-containing precipitate.
- 8. A method for purifying factor XIII from a biological fluid, comprising the steps of:
- a) fractionating a biological fluid by anion exchange chromatography to produce a fraction enriched for factor XIII;
- b) adding Na-acetate to the enriched fraction to form a crystalline precipitate;
- c) dissolving the precipitate to form a solution;
- d) fractionating the solution by anion exchange chromatography to produce a second fraction enriched for factor XIII;
- e) fractionating the second enriched fraction by hydrophobic interaction chromatography to produce a third fraction enriched for factor XIII;
- f) fractionating the third enriched fraction by hydrophobic interaction chromatography to produce a fourth fraction enriched for factor XIII; and
- g) fractionating the fourth enriched fraction by gel filtration and collecting a factor XIII-containing peak fraction.
- 9. A composition of matter comprising factor XIII which is at least 99% pure with respect to contaminating proteins and wherein 1% or less of said factor XIII is factor XIIIa.
- 10. The composition of claim 9 wherein said factor XIII is recombinant human factor XIII.
- 11. The composition of claim 10 wherein said recombinant human factor XIII is yeast-produced recombinant human factor XIII.
- 12. The composition of claim 11 wherein said composition contains less than 50 ppm of yeast protein.
- 13. The composition of claim 11 wherein said composition contains less than 20 ppm of yeast protein.
- 14. The composition of claim 11 wherein said composition contains less than 10 ppm of yeast protein.
- 15. The composition of claim 11 wherein said composition contains less than 1 ppm of yeast protein.
- 16. The composition of claim 15 wherein 0.5% or less of said factor XIII is factor XIIIa.
- 17. The composition of claim 9, wherein 0.5% or less of said factor XIII is factor XIIIa.
- 18. The composition of claim 9, wherein said factor XIII is human factor XIII.
- 19. The composition of claim 9, wherein said factor XIII is a dimer of a subunits.
- 20. A composition of matter comprising yeast-produced recombinant human factor XIII, wherein said composition contains less than 100 ppm yeast protein and wherein 1% or less of said factor XIII is factor XIIIa.
- 21. The composition of claim 20 wherein said composition contains less than 10 ppm yeast protein.
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. patent application Ser. No. 07/927,196, filed Aug. 7, 1992, now abandoned, which is a continuation-in-part of Ser. No. 07/741,263, filed Aug. 7, 1991, now abandoned, which is a continuation-in-part of Ser. No. 07/270,714, filed Nov. 14, 1988, now U.S. Pat. No. 5,204,447, and of Ser. No. 07/525,556, filed May 18, 1990, which application has been abandoned and is a continuation-in-part of U.S. Ser. No. 07/521,805, filed May 10, 1990 which has been abandoned.
US Referenced Citations (5)
Foreign Referenced Citations (1)
Number |
Date |
Country |
6989687 |
Sep 1987 |
AUX |
Related Publications (2)
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Number |
Date |
Country |
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525556 |
May 1990 |
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521805 |
May 1990 |
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Continuations (1)
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Number |
Date |
Country |
Parent |
927196 |
Aug 1992 |
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Continuation in Parts (2)
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Number |
Date |
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Parent |
741263 |
Aug 1991 |
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Parent |
270714 |
Nov 1988 |
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