| 两种凝血酶原提取方法的比较 |
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作者：赖翼 吴民泸 代娟 陈曼 段佳慧 刘阳
【摘要】 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。
【关键词】 凝血酶原； 凝血酶； 等电点沉淀法； 柠檬酸钡吸附法
【Abstract】 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 Ca2+ 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 Ca2+ from prothrombin purified by method of precipitating at isoelectricpoint was 11.01±1.54 U/mg.The specific activity of thrombin activated by Ca2+ from prothrombin purified by adsorption method with barium citrate was 130.17±12.30 U/mg.The specific activities were analyzed by paired Ttest,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.
【Key words】 prothrombin； thrombin； method of precipitating at isoelectricpoint； adsorption method with barium citrate
凝血酶是参与机体凝血过程的一个重要的凝血因子，在体内以凝血酶原形式存在［1］。20世纪中期开始人们将凝血酶应用于临床，由于其疗效显著，应用也越来越广泛。之后美英日等国从牛血浆中分离出凝血酶应用于临床，并证明将动物凝血酶应用于人体未出现凝血酶抗原性，口服和局部外用时无过敏反应和其他不良反应等，因而目前所使用的凝血酶制剂大多来源于动物血浆，国内采用猪血浆制备凝血酶的报道较多［2］。等电点沉淀法和柠檬酸钡吸附法是两种主要的凝血酶原提取方法［3］，凝血酶 [1] [2] [3] 下一页 |
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