Factor VIII Fractionation on Aminohexyl Sepharose with Possible Reduction in Hepatitis B Antigen

Summary

The method of factor VIII purification by chromatography on aminohexyl Sepharose has been extended so that up to 100 ml of intermediate purity concentrate can be processed on an 8.6 ml column. The product has a specific activity of 1.8 International Units of factor VIII per mg of protein. Most of the fibrinogen is removed and antibodies to blood group substances A and B are not detectable by haemagglutination techniques. Hepatitis B antigen has been reduced to one sixteenth of that in the starting material, in preliminary experiments. The process has the added advantage that it concentrates the factor VIII relative to the starting material.

Keywords

Factor VIII - Hepatitis B - Factor VIII fractionation - Aminoheryl Sepharose

• References

• 1 Austen DEG. The chromatographic separation of factor VIII on aminohexyl Sepharose. Br J Haematol 1979; 43: 669-674
• 2 Austen DEG, Rhymes IR. A Laboratory Manual of Blood Coagulation. Blackwell Scientific Publications; Oxford England: 1975: 50-54
• 3 Biggs RP, Eveling J, Richards G. The assay of antihaemophilic-globulin activity. Br J Haematol 1955; 1: 20-34
• 4 Zimmerman TS, Ratnoff OD, Powell AE. Immunologic differentiation of classic haemophilia (factor VIII deficiency) and von Wille-brand's disease. J Clin Invest 1971; 50: 244-254
• 5 Cameron CH. Experiences with solid-phase radioimmunoassay in the rapid diagnosis of hepatitis B. J Med Virol 1978; 3: 27-30
• 6 Wiseman IC. A modification of Hepatest using the Terasaki plate for the detection of HB_sAg in blood donors. J Clin Pathol 1976; 29: 264-268