Phan Jennifer, Yamout Nasrin, Schmidberger Jason, Bottomley Stephen P, Buckle Ashley M
Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2011;752:45-57. doi: 10.1007/978-1-60327-223-0_4.
The expression and harvesting of proteins from insoluble inclusion bodies by solubilization and refolding is a technique commonly used in the production of recombinant proteins. Despite the importance of refolding, publications in the literature are essentially ad hoc reports consisting of a dazzling array of experimental protocols and a diverse collection of buffer cocktails. For the protein scientists, using this information to refold their protein of interest presents enormous challenges. Here, we describe some of the practical considerations in refolding and present several standard protocols. Further, we describe how refolding procedures can be designed and modified using the information in the REFOLD database (http://refold.med.monash.edu.au), a freely available, open repository for protocols describing the refolding and purification of recombinant proteins.
通过溶解和重折叠从不溶性包涵体中表达和收获蛋白质是重组蛋白生产中常用的技术。尽管重折叠很重要，但文献中的出版物基本上都是临时报告，包含一系列令人眼花缭乱的实验方案和各种各样的缓冲液配方。对于蛋白质科学家来说，利用这些信息对他们感兴趣的蛋白质进行重折叠面临着巨大挑战。在这里，我们描述了重折叠中的一些实际考虑因素，并给出了几个标准方案。此外，我们还描述了如何利用REFOLD数据库（http://refold.med.monash.edu.au）中的信息来设计和修改重折叠程序，该数据库是一个免费的开放资源库，用于描述重组蛋白重折叠和纯化的方案。
