Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS

被引：92

作者：

Tao, Hu ^{[1
]}

Liu, Wenjun ^{[1
]}

Simmons, Brandi N. ^{[1
]}

Harris, Helen K. ^{[1
]}

Cox, Timothy C. ^{[2
]}

Massiah, Michael A. ^{[1
]}

机构：

[1] Oklahoma State Univ, Dept Biochem & Mol Biol, Stillwater, OK 74078 USA

[2] Univ Washington, Dept Pediat, Div Craniofacial Med, Seattle, WA 98195 USA

来源：

BIOTECHNIQUES | 2010年 / 48卷 / 01期

基金：

美国国家科学基金会;

关键词：

protein solubilization; inclusion bodies; protein purification; sarkosyl; detergents; MALTOSE-BINDING-PROTEIN; ESCHERICHIA-COLI; FUSION TECHNOLOGY; SOLUBLE-PROTEIN; PURIFICATION; EXPRESSION; POLYPEPTIDES; SOLUBILITY; SUMO;

D O I：

10.2144/000113304

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a rapid, simple, and efficient method for recovering glutathione S-transferase (GST)- and His6-tagged maltose binding protein (MBP) fusion proteins from inclusion bodies. Incubation of inclusion bodies with 10% sarkosyl effectively solubilized >95% of proteins, while high-yield recovery of sarkosyl-solubilized fusion proteins was obtained with a specific ratio of Triton X-100 and CHAPS. We demonstrate for the first time that this combination of three detergents significantly improves binding efficiency of GST and GST fusion proteins to gluthathione (GSH) Sepharose.

引用

页码：61 / 64

页数：3

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