Reduction of the content of protein contaminants in compositions comprising a vitamin K-dependent protein of interest
The present invention relates to Vitamin K-dependent protein compositions having a very low, or negligible, content of protein contaminants. The present invention also relates to method applicable in the preparation of such Vitamin K-dependent protein compositions. Such methods may either be used alone or in sequential combination with the purpose of reducing the relative content of protein contaminants. The present invention is particularly relevant in the preparation of compositions of coagulation factors selected from Factor X polypeptides (FX/FXa), Factor IX polypeptides (FIX/FIXa), Factor VII polypeptides (FVII/FVIIa), and the anticoagulant Protein C, in particular Factor VII polypeptides.
1. A method for reducing the concentration of Protein S in a composition comprising a recombinant Vitamin K-dependent protein of interest produced under cell culturing conditions, the method comprising:
(A) contacting a cell culture supernatant composition comprising a recombinant Vitamin K-dependent protein of interest and Protein S, wherein the recombinant Vitamin K-dependent protein of interest is produced under cell culturing conditions with a chromatographic column comprising a solid phase material canying monoclonal antibodies against the Vitamin K-dependent protein of interest, wherein the Protein S is present in the composition at an initial concentration of 500 to 1000 parts per million (ppm) relative to the Vitamin K-dependent protein of interest;
(B) collecting a resulting composition comprising the Vitamin-K dependent protein of interest and Protein S;
(C) contacting the resulting composition with a chromatographic column comprising a solid phase material carrying monoclonal antibodies against Protein S, wherein the monoclonal antibodies against Protein S selectively bind Protein S but do not bind the recombinant Vitamin K-dependent protein of interest to the solid phase material thereby allowing the Vitamin K-dependent protein of interest to flow through the chromatographic column; and
(D) collecting the resulting composition comprising the Vitamin-K dependent protein of interest and Protein S, wherein the concentration of Protein S in the resulting composition expressed as parts per million relative to the recombinant Vitamin K-dependent protein of interest is reduced to less than or equal to 100 ppm.
2. The method according to claim 1 , wherein the recombinant Vitamin K-dependent protein of interest is human Factor VIIa.