Claims
- 1. A modified human factor VIII comprising an amino acid substitution at one or more of positions 484-508 according to SEQ ID NO:2, said substitution being an insertion of an immunoreactivity-reducing amino acid for the naturally-occurring amino acid, said modified factor VIII having procoagulant activity.
- 2. A modified factor VIII according to claim 1 wherein the modified factor VIII has reduced reactivity to an inhibitory antibody as compared to unmodified factor VIII.
- 3. The modified factor VIII of claim 1 wherein the amino acid substitution is made at one or more of positions selected from the group consisting of 490, 493, 496, 499, 500, 502, 503, 505 and 507.
- 4. The modified factor VIII of claim 1 wherein the amino acid substitutions is made at one or more of positions selected from the group consisting of 486, 491, 494, 498, 504 and 506.
- 5. The modified factor VIII of claim 1 wherein an amino acid substitution is made at position 497.
- 6. The modified factor VIII of claim 1 wherein an amino acid substitution is made at position 490.
- 7. DNA encoding modified human factor VIII A2 domain, said DNA having one or more nucleotide substitutions resulting in a coding change at one or more amino acid positions chosen from positions 484-508 according to SEQ ID NO:2, said change encoding an immunoreactivity-reducing amino acid at the chosen position.
- 8. DNA according to claim 7 wherein the amino acid position chosen is selected from the group consisting of 490, 493, 496, 499, 500, 502, 503, 505 and 507.
- 9. DNA according to claim 7 wherein the amino acid position chosen is selected from the group consisting of 486, 491, 494, 498, 504 and 506.
- 10. DNA according to claim 7 wherein the amino acid position chosen is amino acid position 497.
- 11. DNA according to claim 7 wherein the amino acid position chosen is amino acid position 490.
- 12. Expression product of DNA encoding human or hybrid human/mammalian factor VIII, said DNA comprising DNA encoding a modified A2 domain, the DNA having one or more nucleotide substitutions resulting in a coding change at one or more of amino acid positions chosen from positions 484-508 according to SEQ ID NO:2, said change encoding an immunoreactivity-reducing amino acid at the chosen position. occurring amino acid.
- 13. The expression product of claim 12 wherein the DNA encoding the modified A2 domain encodes an amino acid substitution at one or more of amino acid positions selected from the group consisting of 490, 493, 496, 499, 500, 502, 503, 505 and 507.
- 14. The expression product of claim 12 wherein the DNA encoding the modified A2 domain encodes an amino acid substitution at one or more of amino acid positions selected from the group consisting of 486, 491, 494, 498, 504 and 506.
- 15. The expression product of claim 12 wherein the DNA encoding the modified A2 domain encodes an amino acid substitution at position 497.
- 16. The expression product of claim 12 wherein the DNA encoding the modified A2 domain encodes an amino acid substitution at position 490.
- 17. The method of making a modified mammalian factor VIII A2 domain comprising the steps of mutating DNA encoding the domain at one or more codons encoding amino acids at positions corresponding to 484-508 of human factor VIII whereby one or more mutated codons encoding an amino acid is substituted for a corresponding naturally-occurring codon, and expressing in a host cell DNA comprising the mutated codons, either independently or in contiguous translatable sequence with DNA encoding another domain of factor VIII, whereby a modified factor VIII A2 domain is made.
- 18. The method of claim 17 wherein the DNA to be mutated is human DNA.
- 19. The method of claim 17 wherein the DNA to be mutated is porcine DNA.
- 20. The method of claim 17 wherein the DNA to be mutated is murine DNA.
- 21. The method of claim 17 wherein a mutated codon encodes an immunoreactivity-reducing amino acid.
- 22. The method of claim 21 wherein a mutated codon encodes an amino acid at a position selected from the group consisting of 490, 493, 496, 499, 500, 502, 503, 505 and 507.
- 23. The method of claim 21 wherein a mutated codon encodes an amino acid at a position selected from the group consisting of 486, 491, 494, 498, 504 and 506.
- 24. The method of claim 21 wherein a mutated codon encodes an amino acid at position 497.
- 25. The method of claim 21 wherein a mutated codon encodes an amino acid at position 490.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. patent application Ser. No. 09/315,179 filed May 20, 1999 and claims priority from U.S. patent application Ser. No. 09/037,601 filed Mar. 10, 1998; which is a continuation-in-part of U.S. patent application Ser. No. 08/670,707 filed Jun. 26, 1996, which issued as U.S. Pat. No. 5,859,204, and of International Patent Application No. PCT/US97/11155 filed Jun. 26, 1997. Application Ser. No. 08/670,707 is a continuation-in-part of PCT/US94/13200 entitled “Hybrid Human/Animal Factor VIII” filed Nov. 15, 1994, and of U.S. Ser. No. 08/212,133 entitled “Hybrid Human/Animal Factor VIII” filed Mar. 11, 1994, which is a continuation-in-part of U.S. Ser. No. 07/864,004 entitled “Hybrid Human/Porcine Factor VIII” filed Apr. 7, 1992, and which issued as U.S. Pat. No. 5,364,771 on Nov. 15, 1994. Application No. PCT/US97/11155 is a continuation-in-part of application Ser. No. 08/670,707.
ACKNOWLEDGEMENT OF FEDERAL RESEARCH SUPPORT
[0002] The government has rights in this invention arising from National Institutes of Health Grant No. HL46215 that partially funded the research leading to this invention.
Divisions (1)
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Number |
Date |
Country |
Parent |
09315179 |
May 1999 |
US |
Child |
10131510 |
Apr 2002 |
US |
Continuation in Parts (6)
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Number |
Date |
Country |
Parent |
09037601 |
Mar 1998 |
US |
Child |
09315179 |
May 1999 |
US |
Parent |
08670707 |
Jun 1996 |
US |
Child |
09037601 |
Mar 1998 |
US |
Parent |
PCT/US94/13200 |
Nov 1994 |
US |
Child |
08670707 |
Jun 1996 |
US |
Parent |
08212133 |
Mar 1994 |
US |
Child |
08670707 |
Jun 1996 |
US |
Parent |
07864004 |
Apr 1992 |
US |
Child |
08212133 |
Mar 1994 |
US |
Parent |
PCT/US97/11155 |
Jun 1997 |
US |
Child |
08212133 |
Mar 1994 |
US |