US 5,288,853 A · Factor viii purification process

Abstract

There is provided in accordance with the practice of this invention a process for separating Factor VIII complex from an impure protein fraction containing Factor VIII complex. An aqueous solution of the impure protein fraction containing Factor VIII complex is applied to a heparin-coupled chromatographic medium, to bind the Factor VIII complex to the medium. The Factor VIII is then recovered from the heparin-coupled chromatographic medium by elution with an aqueous solution comprising CaCl.sub.2 and histidine. The recovered Factor VIII is further purified by precipitation with a solution comprising glycine and NaCl, and washing the resultant precipitate with a solution comprising histidine, glycine, and NaCl to provide a Factor VIII complex solution with a specific activity of about 70 to about 150 units/mg.

Claims21

Dependent claims

A process for separating Factor VIII complex from an impure protein fraction containing Factor VIII complex, the process comprising the steps of:

providing an aqueous solution of an impure protein fraction comprising Factor VIII complex;
applying the impure protein fraction solution to a heparin-coupled chromatographic medium to thereby bind Factor VIII complex to the heparin;
eluting the Factor VIII complex from the chromatographic medium using an aqueous solution comprising CaCl.sub.2 ;
adding a sufficient amount of glycine and sodium chloride to the eluate to thereby precipitate Factor VIII complex;
washing the Factor VIII complex precipitate with a wash solution;
and
recovering the washed Factor VIII complex.

4 more elements

Dependencies are inferred from the claim text.

Factor viii purification process

Life of this patent

Factor viii purification process

Life of this patent

Abstract

Claims21