人纤维结合蛋白制备方法的建立及人纤维结合蛋白稳定性初探  

Establishment of a method for human fibronectin preparation and primary study on fibronectin stability

作　　者：祝慈芳[1] 朱威[1] 

机构地区：[1]上海生物制品研究所有限责任公司第五研究室,200052

出　　处：《国际生物制品学杂志》2013年第4期185-189,共5页International Journal of Biologicals

摘　　要：目的 建立人纤维结合蛋白（fibronectin,Fn）制备方法,研究不同保护剂对Fn稳定性的影响.方法 将经磷酸盐缓冲液抽提溶解的血浆冷沉淀用聚乙二醇沉淀,获得的上清液经离子交换层析制得纯化的Fn.将加入复合保护剂的Fn溶液分别于-20、4和37℃存放0、5和14 d,观察保护剂抑制Fn降解的效果.结果 采用建立的方法制备的Fn的纯度和平均回收率均＞80％.Fn稳定性研究显示,Fn于4℃存放会发生降解,加入枸橼酸钠＋甘氨酸或枸橼酸钠＋咪唑复合保护剂能有效抑制Fn降解.结论 采用建立的方法制备Fn是可行的.枸橼酸钠＋甘氨酸或枸橼酸钠＋咪唑复合保护剂对Fn具有保护作用.Objective To establish a method for human fibronectin preparation and study the effect of different stabilizing agents on stability of human fibronectin. Methods The plasma eryoprecipitate dissolved by phosphate buffer was precipitated with polyethylene glycol. The obtained supernatant was purified by ion exchange chromatography to prepare the pure fibronectin. The fibronectin solutions added different compound stabilizing agents were respectively stored at - 20, 4 and 37℃ for 0, 5 and 14 days. The inhibition of different compound stabilizing agents were observed. Results The purity and the average recovery of fibroneetin prepared by the established method were both 〉 80%. The study on stability of fibroneetin showed that fibronectin stored at 4℃ could degrade. Adding the compound stabilizing agent comprised by glycine or imidazole into the fibronectin solutions could effectively inhibit fibronectin degradation at 4℃. Conclusions Using the established method to prepare fibroneetin is available. The compound stabilizing agent comprised by glycine or imidazole can stabilize fibronectin.

关 键 词：纤连蛋白类 色谱法 离子交换 蛋白质稳定性 辅药 

分 类 号：R914[医药卫生—药物化学]