Open AccessOverview of the Purification of Recombinant Proteins
Author(s) -
Wingfield Paul T.
Publication year - 2015
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps0601s80
Subject(s) - protein purification , recombinant dna , flag tag , affinity chromatography , fusion protein , protein expression , computational biology , protein tag , tandem affinity purification , target protein , chemistry , biology , biochemistry , gene , enzyme
When the first version of this unit was written in 1995, protein purification of recombinant proteins was based on a variety of standard chromatographic methods and approaches, many of which were described and mentioned throughout Current Protocols in Protein Science . In the interim, there has been a shift toward an almost universal usage of the affinity or fusion tag. This may not be the case for biotechnology manufacture where affinity tags can complicate producing proteins under regulatory conditions. Regardless of the protein expression system, questions are asked as to which and how many affinity tags to use, where to attach them in the protein, and whether to engineer a self‐cleavage system or simply leave them on. We will briefly address some of these issues. Also, although this overview focuses on E.coli , protein expression and purification, other commonly used expression systems are mentioned and, apart from cell‐breakage methods, protein purification methods and strategies are essentially the same. © 2015 by John Wiley & Sons, Inc.