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Journal of Analytical Methods in Chemistry
Volume 2013 (2013), Article ID 581093, 8 pages
http://dx.doi.org/10.1155/2013/581093
Review Article

Several Affinity Tags Commonly Used in Chromatographic Purification

State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, No. 17, Third Section of Renmin South Road, Chengdu 610041, China

Received 12 July 2013; Revised 11 November 2013; Accepted 2 December 2013

Academic Editor: Kiran Kumar Doddapaneni

Copyright © 2013 Xinyu Zhao et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Affinity tags have become powerful tools from basic biological research to structural and functional proteomics. They were widely used to facilitate the purification and detection of proteins of interest, as well as the separation of protein complexes. Here, we mainly discuss the benefits and drawbacks of several affinity or epitope tags frequently used, including hexahistidine tag, FLAG tag, Strep II tag, streptavidin-binding peptide (SBP) tag, calmodulin-binding peptide (CBP), glutathione S-transferase (GST), maltose-binding protein (MBP), S-tag, HA tag, and c-Myc tag. In some cases, a large-size affinity tag, such as GST or MBP, can significantly impact on the structure and biological activity of the fusion partner protein. So it is usually necessary to excise the tag by protease. The most commonly used endopeptidases are enterokinase, factor Xa, thrombin, tobacco etch virus, and human rhinovirus 3C protease. The proteolysis features of these proteases are described in order to provide a general guidance on the proteolytic removal of the affinity tags.