Variant of antihemophilic factor VIII having increased specific activity
View Patent ↗The present invention is in the field of hemophilia therapy. It relates to a new variant of antihemophilic factor VIII having increased specific activity in comparison to known factor VIII products.
1. A polynucleotide encoding an isolated and recombinant, fully or partially B-domain deleted porcine factor VIII (FVIII) variant, the FVIII variant being devoid of an amino acid sequence corresponding to amino acids 714 to 740 of SEQ ID NO: 7.
2. An expression vector comprising a polynucleotide according to claim 1 .
3. A mammalian cell comprising an expression vector according to claim 2 .
4. A method for producing a FVIII variant, comprising the steps of:
a. Culturing a mammalian cell according to claim 3 ; and
b. Isolating from the mammalian cell the FVIII variant.
5. The method according to claim 4 , further comprising the step of
c. Formulating the factor VIII variant together with appropriate excipients into a pharmaceutical composition.
6. The polynucleotide of claim 1 , wherein the FVIII variant comprises a sequence at least 90% identical to SEQ ID NO: 1, the variant being devoid of the amino acid sequence corresponding to amino acids 714 to 740 of SEQ ID NO: 7.
7. The polynucleotide of claim 6 , wherein the FVIII variant shares at least 95% identity to SEQ ID NO: 1.
8. The polynucleotide of claim 7 , wherein the FVIII variant shares at least 99.5% identity to SEQ ID NO: 1.
9. The polynucleotide of claim 8 , wherein the FVIII variant is SEQ ID NO: 1.
10. The polynucleotide of claim 1 , wherein the FVIII variant is partially B-domain deleted and the remaining portion of the B-domain is selected from a sequence consisting of SEQ ID NO: 4.
11. An expression vector comprising a polynucleotide according to claim 6 .
12. An expression vector comprising a polynucleotide according to claim 7 .
13. An expression vector comprising a polynucleotide according to claim 8 .
14. An expression vector comprising a polynucleotide according to claim 9 .
15. A mammalian cell comprising an expression vector according to claim 11 .
16. A mammalian cell comprising an expression vector according to claim 12 .
17. A mammalian cell comprising an expression vector according to claim 13 .
18. A mammalian cell comprising an expression vector according to claim 14 .
19. A method for producing a FVIII variant, comprising the steps of:
a. Culturing a mammalian cell according to claim 16 ; and
b. Isolating from the mammalian cell the FVIII variant.
20. A method for producing a FVIII variant, comprising the steps of:
a. Culturing a mammalian cell according to claim 18 ; and
b. Isolating from the mammalian cell the FVIII variant.