重组类胰岛素样生长因子-Ⅰ的纯化与复性  

Purification and Refolding of Recombinant Human Insulin-like Growth Factor I

作　　者：刘侃[1] 汪炬[1] 谢秋玲[1] 李志英[1] 洪岸[1] 

机构地区：[1]暨南大学生物工程研究所,广州510632

出　　处：《中国生物工程杂志》2006年第2期29-33,共5页China Biotechnology

基　　金：广东省生物工程药物重点实验室项目资助(2002B601983)

摘　　要：目的:获得高纯度和高活性的胰岛素样生长因子(insulin-like growth factor,IGF-1);方法:构建好的BL21大肠杆菌工程菌经IPTG诱导,以融合一段截短型半乳糖苷酶及His-tag形式表达IGF-1融合蛋白(约15kDa),超声破碎,提取包涵体经镍柱亲和层析后,用羟氨切割纯化的融合蛋白,纯化后的蛋白质在小分子保护剂及GSH/GSSG的存在下复性。结果:经Ni2+柱亲和层析,IGF-1纯度达90%以上,复性后得到有较高生物活性的IGF-1。结论:IGF-1发酵及纯化和复性方法的建立为大量生产IGF-1打下了基础。Objective： To obtain Insulin-like growth factor （IGF-1） with high purity and activity; Methods： His-tag-beta-galactosidase-IGF-1 fusion protein was expressed in Escherichia colt as inclusion body with IPTG induction. Cells were then harvested, sonicated and centrifuged , and the inclusion bodies were isolated and purified by Ni^2＋-high performance affinity chromatography. After cleavage of the fusion protein with hydroxylamine, the released IGF-1 was purified by Ni^2＋- high performance affinity chromatography again and refolded in the presence of GSH/GSSH. Results： The purity of the released IGF-1 was more than 90% after Ni^2＋-high performance affinity chromatography, and the refolded IGF-1 was with high biological activities. Conclusion： The procudure of fermentation, simple purification and renaturation of recombinant IGF-1 could build the foundation for the large-scale production of IGF-1.

关 键 词：胰岛素样生长因子-1(IGF-1) 纯化复性 

分 类 号：Q959.468[生物学—动物学] S852.21[农业科学—基础兽医学]