Factor VII or VIIA-like molecules
Conjugates of Factor VII (FVII) and Factor VIIa (FVIIA) are provided, as are methods for preparing them. Methods for producing novel polypeptides contributing to the production of such conjugates are provided. Methods of treatment by administering a FVII or FVIIa conjugate are provided.
1. A nucleic acid comprising a nucleotide sequence which encodes a polypeptide comprising an amino acid sequence which (a) differs from the hFVII or hFVIIa sequence SEQ ID NO:1 in 1-15 amino acid residues and (b) comprises an introduced in vivo N-glycosylation site relative to SEQ ID NO:1, wherein the introduced in vivo N-glycosylation site comprises the substitution V253N.
2. The nucleic acid of claim 1 , wherein the encoded polypeptide further comprises at least one additional introduced in vivo N-glycosylation site.
3. The nucleic acid of claim 2 , wherein the at least one additional introduced in vivo N-glycosylation site comprises the substitution I205S/T.
4. The nucleic acid of claim 2 , wherein the at least one additional introduced in vivo N-glycosylation site comprises the substitutions R315N+V317S/T.
5. The nucleic acid of claim 2 , wherein the encoded polypeptide comprises two additional introduced in vivo N-glycosylation sites comprising the substitutions I205S/T and T267N.
6. The nucleic acid of claim 2 , wherein the additional introduced in vivo N-glycosylation site is introduced in a position relative to SEQ ID NO:1 occupied by an amino acid residue with more than 25% of its side chain exposed to the solvent.
7. The nucleic acid of claim 2 , wherein the additional introduced in vivo N-glycosylation site comprises an amino acid substitution in a position selected from the group consisting of 28-48, 139-147, 286-294, 311-319, 338-345 and 388-406 relative to SEQ ID NO:1.
8. An expression vector comprising the nucleic acid of claim 1 .
9. A host cell comprising the nucleic acid of claim 1 .
10. The host cell of claim 9 , wherein the host cell is a eukaryotic host cell.