Claims
- 1. A modified human factor VIII comprising an amino acid substitution at one or more positions selected from the group consisting of 2199, 2200, 2223, 2227, and 2252.
- 2. The modified human factor VIII of claim 1 lacking a B-domain.
- 3. The modified human factor VIII of claim 1 comprising isoleucine substituted for methionine 2199.
- 4. The modified human factor VIII of claim 1 comprising leucine substituted for phenylalanine 2200.
- 5. The modified human factor VIII of claim 1 comprising phenylalanine substituted for leucine 2252.
- 6. The modified human factor VIII of claim 1 comprising isoleucine substituted for methionine 2199 and leucine substituted for phenylalanine 2200.
- 7. The modified human factor VIII of claim 1 comprising alanine substituted for valine 2223 and glutamate substituted for lysine 2227.
- 8. The modified human factor VIII of claim 1 comprising isoleucine substituted for methionine 2199, leucine substituted for phenylalanine 2200, alanine substituted for valine 2223, and glutamate substituted for lysine 2227.
- 9. The modified factor VIII of claim 1 which has reduced antigenicity as compared to the corresponding human protein.
- 10. The modified factor VIII of claim 1 which has reduced immunogenicity as compared to the corresponding human protein
- 11. The modified factor VIII of claim 1 which has reduced immunogenicity and reduced antigenicity as compared to the corresponding human protein.
- 12. The modified factor VIII of claim 1 which has a specific activity greater than about 2,000 units per milligram.
- 13. The modified factor VIII of claim 1 which has a specific activity greater than about 3,000 units per milligram.
- 14. The modified factor VIII of claim 1 which has a specific activity greater than about 5,000 units per milligram.
- 15. The modified factor VIII of claim 1 which has a specific activity greater than about 10,000 units per milligram.
- 16. The modified factor VIII of claim 1 which is a single mutant.
- 17. The modified factor VIII of claim 1 which is a double mutant.
- 18. The modified factor VIII of claim 1 which is a triple mutant.
- 19. The modified factor VIII of claim 1 which is a quadruple mutant.
- 20. The modified factor VIII of claim 1 which has lower antigenicity towards at least one C2-specific inhibitory antibody as compared to human factor VIII.
- 21. The modified factor VIII of claim 1 which has an increased or decreased Bethesda titer of monoclonal antibody B02011 as compared to human factor VIII or full length recombinant factor VIII.
- 22. The modified factor VIII of claim 1 which has an increased or decreased Bethesda titer of monoclonal antibody NMC VIII-S as compared to human factor VIII or full-length recombinant factor VIII.
- 23. The modified factor VIII of claim 1 which has an increased or decreased Bethesda titer towards at least one inhibitory antibody preparation as compared to human factor VIII or recombinant human factor VIII.
- 24. A modified factor VIII comprising at least one amino acid substitution of a non-human factor VIII amino acid for the corresponding human factor VIII amino acid.
- 25. The modified factor VIII of claim 24 wherein the at least one non-human factor VIII amino acid substitution is from a non-human mammal.
- 26. The modified factor VIII of claim 25 wherein the non-human mammal is porcine, canine and/or murine.
- 27. The modified factor VIII of claim 25 which has coagulant activity and improved antigenicity as compared to the factor VIII molecule from which it was derived or other factor VIII preparations.
- 28. The modified factor VIII of claim 25 wherein the amino acid substitution is not alanine.
- 29. The modified factor VIII of claim 25 which has reduced immunogenicity as compared to the factor VIII molecule from which it was derived or other factor VIII molecules.
- 30. A method for modifying a factor VIII such that reactivity to an inhibitory antibody is reduced and procoagulant activity is retained comprising substituting an immuno-reactivity reducing amino acid for the naturally occurring amino acid at at least one of positions 2199, 2200, 2223, 2227 and 2252.
- 31. The method of claim 30 wherein the at least one substituted amino acid is position 2199.
- 32. The method of claim 30 wherein the at least one substituted amino acid is position 2200.
- 33. The method of claim 30 wherein the at least one substituted amino acid is position 2223.
- 34. The method of claim 30 wherein the at least one substituted amino acid is position 2227.
- 35. The method of claim 30 wherein the at least one substituted amino acid is position 2252.
- 36. The method of claim 30 wherein the modified factor VIII is a single mutant.
- 37. The method of claim 30 wherein the modified factor VIII is a double mutant.
- 38. The method of claim 30 wherein the modified factor VIII is a triple mutant.
- 39. The method of claim 30 wherein the modified factor VIII is a quadruple mutant.
- 40. A method for modifying factor VIII such that antigenicity is reduced and procoagulant activity is retained comprising substituting an immuno-reactivity reducing amino acid for the naturally occurring amino acid at at least one of positions 2199, 2200, 2223, 2227 and 2252.
- 41. The method of claim 40 wherein the at least one substituted amino acid is position 2199.
- 42. The method of claim 40 wherein the at least one substituted amino acid is position 2200.
- 43. The method of claim 40 wherein the at least one substituted amino acid is position 2223.
- 44. The method of claim 40 wherein the at least one substituted amino acid is position 2227.
- 45. The method of claim 40 wherein the at least one substituted amino acid is position 2252.
- 46. The method of claim 40 wherein the modified factor VIII is a single mutant.
- 47. The method of claim 40 wherein the modified factor VIII is a double mutant.
- 48. The method of claim 40 wherein the modified factor VIII is a triple mutant.
- 49. The method of claim 40 wherein the modified factor VIII is a quadruple mutant.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Applications 60/236,460 filed Sep. 29, 2000, and 60/234,047 filed Sep. 19, 2000, both of which are hereby incorporated by reference to the extent not inconsistent with the disclosure herein.
ACKNOWLEDGMENT OF FEDERAL RESEARCH SUPPORT
[0002] This invention was made, at least in part, with funding from the National Institutes of Health under contract No. FO1-HL46215. Accordingly, the U.S. government may have certain rights in this invention.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60236460 |
Sep 2000 |
US |
|
60234047 |
Sep 2000 |
US |