METHOD OF PRODUCING FACTOR VIII PROTEINS BY RECOMBINANT METHODS

摘要

Provided herein are methods and compositions for producing Factor VIII proteins. Such methods include introducing into a cell a nucleic acid molecule encoding a Factor VIII protein operably linked to a promoter, wherein the promoter is characterized by the ability to produce commercially viable Factor VIII protein; and incubating the cell under conditions for producing commercially viable Factor VIII protein. Also provided are nucleic acid molecules which encode a Factor VIII protein operably linked to a Chinese hamster elongation factor 1-a (CHEF1) promoter, which may be used in the methods provided herein.

主权项

1. A method for producing a recombinant Factor VIII protein, comprising the steps of:
a. introducing into a cell a nucleic acid molecule encoding a modified human Factor VIII protein operably linked to a Chinese hamster elongation factor 1-alpha promoter, wherein said promoter is characterized by the ability to produce commercially viable Factor VIII protein and wherein said modification comprises a truncated or deleted B domain, a SQ linker, and a Pace/Furin cleavage site, characterized in that upon Pace/Furin cleavage and secretion the protein becomes a two-chain form comprising an A1-A2 chain and an A3-C1-C2 chain; and b. incubating the cell under appropriate conditions for producing commercially viable Factor VIII protein.