Method of producing recombinant vitamin k dependent proteins
AssigneeCnj Holdings Inc
InventorsWilliam N. Drohan, Michael J. Griffith
Life of this patent
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Abstract
Methods for producing cell lines with high levels of biologically active recombinant vitamin K dependent proteins are described. The transfected cell lines do not include heterologous genes for processing enzymes and are not subject to selection pressure such as methotrexate resistance. Cell lines producing Factor VII/VIIa and Factor IX are described. These cell lines can be used for isolation of Factor VII/VIIa and/or Factor IX for treatment of Hemophilia.
Claims20
A method of producing a vitamin K dependent protein, the method comprising the steps of:
- (a) transfecting a population of mammalian cells with a gene encoding the vitamin K dependent protein operably linked to a promoter;
- (b) performing at least one round of cloning and screening to identify cells which produce at least 10 mg/L of the vitamin K dependent protein, wherein at least 10% of the vitamin K dependent protein is biologically active;
- (c) optionally, repeating step (b) one or more times;
- and
- (d) harvesting the vitamin K dependent protein.
A method of producing a vitamin K dependent protein, the method comprising the steps of:
- (a) transfecting a population of mammalian cells with a gene encoding the vitamin K dependent protein operably linked to a Chinese hamster elongation factor 1 (CHEF-1) promoter;
- and
- (b) performing at least one round of cloning and screening to identify cells which produce at least 10 mg/L of the vitamin K dependent protein, wherein at least 10% of the vitamin K dependent protein is biologically active.
Dependencies are inferred from the claim text.