猪血凝血酶制备工艺

Preparation process of porcine thrombin

分别使用等电点沉淀法和柠檬酸钡吸附法从猪血浆中提取凝血酶原,凝血酶原经单独钙离子激活得到凝血酶,通过凝血酶效价测定、蛋白质浓度测定,计算出凝血酶的比活。通过单因素试验和正交试验,考察激活时间、Ca2+浓度、温度对凝血酶活力的影响。结果表明:与等电点沉淀法相比,采用柠檬酸钡吸附法提取得到的凝血酶原经激活得到的凝血酶比活力高43U/mg;最佳激活条件Ca2+浓度0.1mol/L,温度25℃下激活2h,在该条件下进行得到凝血酶比活力最大。

Prothrombin was purified by method of precipitating at isoelectricpoint and adsorption method with barium citrate from blood plasma of pig and activated by Ca^2＋ only to thrombin.Then the titer of thrombin and the concentration of protein were measured to get the specific activity of thrombin,using the single factor tests and orthogonal experiments to study the activity of thrombin under different conditions.Results：The specific activity of thrombin by adsorption method with barium citrate was 43 U/mg higher than that by method of precipitating at isoelectricpoint;The optimum activating time and the concentration of Ca^2＋ were 2 hours and 0.1 mol/L at 25 ℃.Under the condition thrombin can own the highest activity,so the condition was the optimum condition.