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Abstract

A rapid and simple process for purifying human, bovine and porcine procoagulant protein Factor VIII on a large scale using sequential high performance size exclusion chromatography under, first, low salt concentration conditions and, second, under high salt concentration conditions from reconstituted commercial Factor VIII:C (complexed Factor VIII) concentrate. The chromatographic separation is carried out on a high performance size exclusion chromatographic column packed with porous beads having a particle size of from about 13 to about 35 microns, pore diameters of from about 500 to about 2000 Angstroms and a pore volume of from about 1.0 to about 1.8 ml per gram. The first chromatographic separation is carried in a buffered aqueous solution using the buffered aqueous solution as an eluant. The low molecular weight constituents (impurities) are separated from Factor VIII and the high molecular weight constituents (impurities). A second chromatographic separation may be carried out after Factor VIII has been dissociated in a buffered solution having a concentration of from about 0.25 to about 0.45M calcium ion. The second chromatographic column is packed with some packing as the first column and is eluted with a buffered aqueous solution containing 0.25 to 0.45M calcium ion. In a column of 2.5.times.60 cm, 4 gms of commercial Factor VIII concentrate can be purified in less than two hours. The process is amenable to scale up.

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