Madaras F, Bell W R, Castaldi P A
Haemostasis. 1978;7(6):321-31. doi: 10.1159/000214276.
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.
已开发出一种简单方法，可从血浆中分离出凝血因子VIII（F VIII），其形式能在兔体内诱导产生单特异性抗体。从血浆中进行氨基酸沉淀后，在琼脂糖4B上进行凝胶过滤，通过肝素-琼脂糖亲和层析进一步纯化。如此分离得到的物质经免疫电泳鉴定，且缺乏凝血活性。兔体内产生的抗血清抑制F VIII凝血活性以及用瑞斯托霉素诱导的血小板凝集所测定的血管性血友病因子活性。用溴化氰-琼脂糖4B使其不溶性化的分离IgG组分保留了与F VIII形成复合物的能力。该复合物具有F VIII凝血活性，可在0.6M氯化钠中去除，并在8M尿素中解离。
