两种凝血酶原提取方法的比较

Comparative Analysis of Two Extraction Methods of Prothrombin

目的比较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的纯度。方法分别采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性。结果采用等电点沉淀法提取的凝血酶原经激活后得到的凝血酶比活性为11.01±1.54 U/mg,采用柠檬酸钡吸附法提取的凝血酶原经激活后得到的凝血酶比活性为130.17±12.30 U/mg,经配对T检验,P<0.01.结论与等电点沉淀法相比,采用柠檬酸钡吸附法从猪血浆中提取得到的凝血酶原纯度更高。

Objective To compare the purity of prothrombin purified by method of precipitating at isoelectricpoint and adsorption method with barium citrate from blood plasma of pig. Methods 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. Results The specific activity of thrombin activated by Ca^2＋ from prothrombin purified by method of precipitating at isoelectricpoint was 11.01± 1.54 U/rag. The specific activity of thrombin activated by Ca^2＋ from prothrombin purified by adsorption method with barium citrate was 130. 17± 12.30 U/rag. The specific activities were analyzed by paired T-test, P〈0. 01. Conclusion The purity of prothrombin purified by adsorption method with barium citrate is higher than purified by method of precipitating at isoelectricpoint.