国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2013年
4期
185-189
,共5页
纤连蛋白类%色谱法,离子交换%蛋白质稳定性%辅药
纖連蛋白類%色譜法,離子交換%蛋白質穩定性%輔藥
섬련단백류%색보법,리자교환%단백질은정성%보약
Fibronectin%Chromatography,ion exchonge%Protein stability%Excipients
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.
目的 建立人纖維結閤蛋白(fibronectin,Fn)製備方法,研究不同保護劑對Fn穩定性的影響.方法 將經燐痠鹽緩遲液抽提溶解的血漿冷沉澱用聚乙二醇沉澱,穫得的上清液經離子交換層析製得純化的Fn.將加入複閤保護劑的Fn溶液分彆于-20、4和37℃存放0、5和14 d,觀察保護劑抑製Fn降解的效果.結果 採用建立的方法製備的Fn的純度和平均迴收率均＞80％.Fn穩定性研究顯示,Fn于4℃存放會髮生降解,加入枸櫞痠鈉+甘氨痠或枸櫞痠鈉+咪唑複閤保護劑能有效抑製Fn降解.結論 採用建立的方法製備Fn是可行的.枸櫞痠鈉+甘氨痠或枸櫞痠鈉+咪唑複閤保護劑對Fn具有保護作用.
목적 건립인섬유결합단백(fibronectin,Fn)제비방법,연구불동보호제대Fn은정성적영향.방법 장경린산염완충액추제용해적혈장랭침정용취을이순침정,획득적상청액경리자교환층석제득순화적Fn.장가입복합보호제적Fn용액분별우-20、4화37℃존방0、5화14 d,관찰보호제억제Fn강해적효과.결과 채용건립적방법제비적Fn적순도화평균회수솔균＞80％.Fn은정성연구현시,Fn우4℃존방회발생강해,가입구연산납+감안산혹구연산납+미서복합보호제능유효억제Fn강해.결론 채용건립적방법제비Fn시가행적.구연산납+감안산혹구연산납+미서복합보호제대Fn구유보호작용.
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 cryoprecipitate 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 fibronectin prepared by the established method were both ＞ 80％.The study on stability of fibronectin 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 fibronectin is available.The compound stabilizing agent comprised by glycine or imidazole can stabilize fibronectin.
