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在高滴度HBsAg混和血浆中加入氯化钙和凝血酶,使其转化成血清后,分别用5种方法提纯,并比较不同方法的提纯效果。方法1:HBsAg经两次离心沉淀(105000×g)后,用超声处理悬浮沉淀物,调至pH2.4。在氯化铯中进行两次等密度超离心,最后用蔗糖作速率区带超离心。方法2:用8%聚乙二醇(PEG)沉淀血清蛋白后,上清液中再加入16%的PEG使HBsAg沉淀。再悬浮后用胃蛋白酶处理。超滤浓缩后作柱层析。方法3:用饱和硫酸铵(30%)沉淀血清
Calcium chloride and thrombin were added to high titer HBsAg mixed plasma to make them into serum, then purified by five kinds of methods respectively, and compared with different purification methods. Method 1: After two centrifugation (105000 × g) of HBsAg, the suspended precipitate was sonicated and adjusted to pH 2.4. Two equal density ultracentrifugations were performed in cesium chloride and finally sucrose was used as a rate zone for ultracentrifugation. Method 2: After precipitating serum proteins with 8% polyethylene glycol (PEG), additional 16% PEG in the supernatant precipitated HBsAg. After resuspension with pepsin treatment. After concentration by ultrafiltration column chromatography. Method 3: Serum was precipitated with saturated ammonium sulfate (30%)