Fully automated protein purification

被引:24
作者
Camper, DeMarco V. [1 ]
Viola, Ronald E. [1 ]
机构
[1] Univ Toledo, Dept Chem, Toledo, OH 43606 USA
基金
美国国家卫生研究院;
关键词
Protein purification; Aspartate semialdehyde dehydrogenase; AKTA Explorer; Automated purification protocols; BETA-SEMIALDEHYDE DEHYDROGENASE; CHELATE AFFINITY-CHROMATOGRAPHY; RECOMBINANT PROTEINS; ESCHERICHIA-COLI; TAGGED-PROTEINS; EXPRESSION; MECHANISM; YIELD;
D O I
10.1016/j.ab.2009.07.009
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Obtaining highly purified proteins is essential to begin investigating their functional and structural properties. The steps that are typically involved in purifying proteins can include an initial capture, intermediate purification, and a final polishing step. Completing these steps can take several days and require frequent attention to ensure success. Our goal was to design automated protocols that would allow the purification of proteins with minimal operator intervention. Separate methods have been produced and tested that automate the sample loading, column washing, sample elution and peak collection steps for ion exchange, metal affinity, hydrophobic interaction, and gel filtration chromatography. These individual methods are designed to be coupled and run sequentially in any order to achieve a flexible and fully automated protein purification protocol. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:176 / 181
页数:6
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