Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
研究报告     
重组类胰岛素样生长因子-Ⅰ的纯化与复性
刘侃 汪炬 谢秋玲 李志英 洪岸
暨南大学生物工程研究所 暨南大学生物工程研究所
Purification and Refolding of Recombinant human insulin-like
 全文: PDF  HTML
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15,000Da),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。
Abstract: 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 coli as inclusion body with IPTG induction. Cells were then harvested, sonicated and centrifuged , and the inclusion bodies were isolated and purified by Ni2+-high performance affinity chromatography. After cleavage of the fusion protein with hydroxylamine, the released IGF-1 was purified by Ni2+- 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 Ni2+-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.
收稿日期: 2005-09-08 出版日期: 2006-02-25
通讯作者: 洪岸   
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
汪炬
李志英
洪岸
谢秋玲
刘侃

引用本文:

刘侃,汪炬,谢秋玲,李志英,洪岸. 重组类胰岛素样生长因子-Ⅰ的纯化与复性[J]. 中国生物工程杂志, .

. Purification and Refolding of Recombinant human insulin-like. China Biotechnology, .

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I02/29

No related articles found!