从血浆冷沉淀中同时制备因子Ⅷ、纤维结合蛋白和纤维蛋白原方法的建立  

Establishment of a method for simultaneous preparation of factor Ⅷ, fibronectin and fibrinogen from plasma cryoprecipitate

作　　者：祝慈芳 朱威 卞炯 

机构地区：[1]生物制品研究所有限责任公司第五研究室,200052上海

出　　处：《国际生物制品学杂志》2013年第5期240-243,247,共5页International Journal of Biologicals

摘　　要：目的 建立从血浆冷沉淀中同时制备因子Ⅷ（factorⅧ,FⅧ）、纤维结合蛋白（fibronectin,Fn）和纤维蛋白原（fibrinogen,Fg）的方法.方法 用缓冲液溶解抽提冷沉淀,获得的冷沉淀抽提液通过2次DEAE离子交换层析和1次聚乙二醇沉淀进行分离纯化来同时制备FⅧ、Fn和Fg.采用FⅧ∶C活性测定试剂盒检测制备的FⅧ活性,并计算FⅧ比活和回收率.分别用双向和单向免疫扩散试验对制备的Fn和Fg进行定性和定量检测,并计算回收率.结果 制备的FⅧ的平均活性为16.48 U/ml,平均比活为29.84 U/mg,回收率＞60％.制备的Fn和Fg的蛋白纯度均＞90％,回收率分别为＞80％和＞75％.结论 采用建立的方法可从冷沉淀中同时制备FⅧ、Fn和Fg.Objective To establish a method for simultaneous preparation of factor Ⅷ （FⅧ）,fibronectin （Fn） and fibrinogen （Fg） from plasma cryoprecipitate.Methods The plasma cryoprecipitate was dissolved and extracted by buffer.The cryoprecipitate solution was separated and purified by twice DEAE ion exchange chromatography and once polyethylene glycol precipitation to prepare FⅧ,Fn and Fg simultaneously.FⅧ：C activity of FⅧ was detected by a detection kit.Specific activity and recovery of FⅧ were calculated.Qualities and quantities of Fn and Fg were detected respectively by double and single immunodiffusion tests.Recoveries of Fn and Fg were calculated.Results F Ⅷ：C activity,specific activity and recovery of the prepared FⅧ were 16.48 U/ml,29.84 U/mg and ＞60％,respectively.Protein purities of the prepared Fn and Fg were both ＞90％,and recoveries of Fn and Fg were ＞ 80％ and ＞ 75％ respectively.Conclusion FⅧ,Fn and Fg can be prepared simultaneously from plasma cryoprecipitate by the established method.

关 键 词：因子Ⅷ 纤连蛋白类 纤维蛋白原 色谱法 离子交换 

分 类 号：R944[医药卫生—药剂学]