Isolation and insolubilisation of human F VIII by affinity chromatography.
A simple procedure has been developed for the isolation of coagulation factor VIII (F VIII) from plasma in a form which induces a monospecific antibody in rabbits. Amino acid precipitation from plasma was followed by gel filtration on Sepharose 4B and further purification was achieved by affinity chromatography on heparin-Sepharose. The material so isolated was identified by immunoelectrophoresis and lacked coagulant activity. The antiserum produced in rabbits inhibited F VIII coagulant activity and von Willebrand factor activity as measured by ristocetin platelet agglutination. The isolated IgG fraction insolubilised with CNBr-Sepharose 4B retained the ability to complex with F VIII. This complex possessed F VIII coagulant activity which could be removed in 0.6 M NaCl and dissociated in 8 M urea.
D000596:Amino Acids; D000918:Antibody Specificity; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D004355:Drug Stability; D005169:Factor VIII / Q000302:isolation & purification*; D006493:Heparin; D006801:Humans; D012685:Sepharose; D012995:Solubility
D000596:Amino Acids; D005169:Factor VIII; D006493:Heparin; D012685:Sepharose