| abstract |
Highly purified, biologically active Human Antihemophilic Factor (AHF) preparations are prepared having specific activities of about 4000 - 8000 units per milligram of AHF. In the method of preparation an AHF concentrate, prepared by fractionation of plasma to partially remove fibrinogen, fibronectin and other plasma components is subjected to a separation on the basis of Stokes' radius to separate AHF from the bulk of remaining proteins in the AHF concentrate. The pooled fractions containing AHF activity are concentrated by precipitation with ammonium sulfate, sodium sulfate, etc., by diafiltration, by PEG addition, or the like. The concentrate, is solubilized or equilibrated in an aqueous medium and treated to change the effective Stokes' radius of the AHF to an apparently low value and then subjected to a separation from the concentrate. The AHF pool from above is treated to remove cations by dialysis against an appropriate buffer of lower ionic strength and chromatographed on an anion-exchange medium. The AHF fraction from the above chromatography, is a highly purified AHF preparation. |