Purify First: Rapid expression and purification of proteins from XMRV

被引:17
|
作者
Gillette, William K. [1 ]
Esposito, Dominic [1 ]
Taylor, Troy E. [1 ]
Hopkins, Ralph F. [1 ]
Bagni, Rachel K. [1 ]
Hartley, James L. [1 ]
机构
[1] NCI Frederick, Prot Express Lab, SAIC Frederick Inc, Frederick, MD 21702 USA
基金
美国国家卫生研究院;
关键词
XMRV; Protein purification; Small scale purification; Parallel purification; VIRUS; SITE;
D O I
10.1016/j.pep.2010.12.003
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Purifying proteins from recombinant sources is often difficult, time-consuming, and costly. We have recently instituted a series of improvements in our protein purification pipeline that allows much more accurate choice of expression host and conditions and purification protocols. The key elements are parallel cloning, small scale parallel expression and lysate preparation, and small scale parallel protein purification. Compared to analyzing expression data only, results from multiple small scale protein purifications predict success at scale-up with greatly improved reliability. Using these new procedures we purified eight of nine proteins from xenotropic murine leukemia virus-related virus (XMRV) on the first attempt at large scale. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:238 / 247
页数:10
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