Current strategies for the use of aReprint of : Y nity tags and tag removal for the puri W cation of recombinant proteins

Content Provider	Semantic Scholar
Author	Arnau, J. M. Lauritzen, Conni Petersen, Gitte Ebert Pedersen, John Kim
Copyright Year	2011
Abstract	AYnity tags are highly eYcient tools for protein puriWcation. They allow the puriWcation of virtually any protein without any prior knowledge of its biochemical properties. The use of aYnity tags has therefore become widespread in several areas of research e.g., high throughput expression studies aimed at Wnding a biological function to large numbers of yet uncharacterized proteins. In some cases, the presence of the aYnity tag in the recombinant protein is unwanted or may represent a disadvantage for the projected application of the protein, like for clinical use. Therefore, an increasing number of approaches are available at present that are designed for the removal of the aYnity tag from the recombinant protein. Most of these methods employ recombinant endoproteases that recognize a speciWc sequence. These process enzymes can subsequently be removed from the process by aYnity puriWcation, since they also include a tag. Here, a survey of the most common aYnity tags and the current methods for tag removal is presented, with special emphasis on the removal of N-terminal histidine tags using TAGZyme, a system based on exopeptidase cleavage. In the quest to reduce the signiWcant costs associated with protein puriWcation at large scale, relevant aspects involved in the development of downstream processes for pharmaceutical protein production that incorporate a tag removal step are also discussed. A comparison of the yield of standard vs. aYnity puriWcation together with an example of tag removal using TAGZyme is also included.
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://wolfson.huji.ac.il/purification/PDF/Literature/Arnau2006.pdf
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article