Process for producing and purifying factor viii and its...

Details Process for producing and purifying factor viii and its... Process for producing and purifying factor viii and its...

C - Chemistry, Metallurgy

12

P

C12P 21/00 (2006.01) C07K 14/755 (2006.01)

Patent

CA 2679107

Disclosed is a method for producing proteins having factor VIII procoagulant activity in serum-free medium by in vitro culturing of mammalian cells, wherein the serum-free medium contains an inhibitor against the protease released from cultured cells. In accordance with this invention, the inhibitor can protect the cleavage of a target protein during cultivation and increase homogeneity of a target molecule, wherein the inhibitor can be a dextran sulfate. This invention also relates to a method of purifying target molecules from the culture medium containing both a target molecule and selected inhibitors by affinity chromatography.

L'invention porte sur une méthode de production de protéines présentant l'activité procoagulante du facteur VIII par culture in vitro de cellules de mammifères dans un milieu sans sérum contenant un inhibiteur agissant contre la protéase libérée par les cellules de la culture. Selon l'invention, l'inhibiteur peut protéger le clivage d'une protéine cible pendant la culture et augmenter l'homogénéité d'une molécule cible. L'inhibiteur peut être du sulfate de dextran. L'invention porte également sur une méthode de purification de molécules cibles d'un milieu de culture contenant à la fois une molécule cible et des inhibiteurs sélectionnés par chromatographie d'affinités.

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