Process for the preparation of human factor VIII and analogs of factor VIII
AssigneeTM Innovation
InventorsGerard Mignot, Nicolas Bihoreau, Philippe Adamowicz
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Abstract
The present invention relates to a process for the preparation of factor VIII or an analog of factor VIII by culture, in a culture medium, of cells which produce said factor VIII or analog of factor VIII and the separation of factor VIII or its analog, wherein the culture medium contains at least one derivative of a polycationic and/or polyanionic polymer. It also relates to factor VIII or analog of factor VIII and to the complex of factor VIII or analog of factor VIII with a derivative of a polycationic and/or polyanionic polymer obtained by using this process.
Claims8
In a process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analog of factor VIII, proviso they are not chosen among:
- Vero, Hela, WI38, BIIK, CHO, COS-7 and MDCK, said mammalian cells expressing the molecule of factor VIII or analog of factor VIII in a continuous mariner, and then separating said factor VIII or analog of factor VIII wherein the improvement comprises culturing the mammalian cells in a serum free culture medium and free of Von Willebrand factor, supplemented with at least one derivative which is a sulfated polysaccharide of molecular weight between 5,000 and 700,000 and a degree of sulfation of 0.5% to 18% by weight of sufur, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said sulfated polysaccharide.
In a process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analog of factor VIII said cells being chosen among recombinant Veto, Hela, WI38, BHK, CHO, COS-7 and MDCK cells which express the molecule of factor VIII or analog of factor VIII in a continuous manner, and then separating said factor VIII or analog of factor VIII, wherein the improvement comprises culturing the mammalian cells in a serum free culture medium and free of Von Willebrand factor, supplemented with at least one derivative which is a sulfated polysaccharide of molecular weight between 5,000 and 700,000 and a degree of sulfation of 65% to 18% by weight of sulfur, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said sulfated polysaccharide proviso that said sulfated polysaccharide is not chosen among heparin, sulfated dextran and hydroxyethylstarch sulfate of molecular weight between 5000 and 700,000 and a degree of sulfation of 0.5% to 18% by weight of sulfur.
A process for the preparation and separation of factor VIII or an analog of factor VIII, the process comprising culturing mammalian eukaryotic cells which produce and secrete said factor VIII or analog of factor VIII, said mammalian eukaryotic cells having incorporated an expression vector containing a gene encoding for said factor VIII or analogue of factor VIII, proviso though they are not chosen among:
- Vero, Hela, WI38, BIIK, CHO, COS-7, MDCK, said mammalian cells expressing the molecule factor VIII or analog of factor VIII in a continuous manner, and then separating said factor VIII or analog of factor VIII wherein the improvement comprises culturing the mammalian cells in a serum free culture median and free of Von Willebrand factor, supplemented with a sulfated polysaccharide selected from the group consisting of heparin, sulfated dextran, and hydroxyethyl starch, in an amount sufficient to provide an enhancement of factor VIII or analog of factor VIII production compared with culturing the mammalian cells in a medium without said supplement.
Dependencies are inferred from the claim text.