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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

  • 祝慈芳 朱威 卞炯
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  • 200052 上海生物制品研究所有限责任公司第五研究室

网络出版日期: 2025-08-16

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

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  • No.5 Research Laboratory, Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China

Online published: 2025-08-16

摘要

目的  建立从血浆冷沉淀中同时制备因子Ⅷ(factor Ⅷ,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。

本文引用格式

祝慈芳 朱威 卞炯 . 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[J]. 国际生物制品学杂志, 2013 , 36(5) : 240 -243,247 . DOI: 10.3760/cma.j.issn.1673-4211.2013.05.005

Abstract

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  Average FⅧ∶C activity, average 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.
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