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.
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. Tn class="Chemical">his 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.
Authors: A M Mildner; D J Rothrock; J W Leone; C A Bannow; J M Lull; I M Reardon; J L Sarcich; W J Howe; C S Tomich; C W Smith Journal: Biochemistry Date: 1994-08-16 Impact factor: 3.162
Authors: Christina J Welch; Melanie L Talaga; Priyanka D Kadav; Jared L Edwards; Purnima Bandyopadhyay; Tarun K Dam Journal: J Biol Chem Date: 2019-12-02 Impact factor: 5.157
Authors: Susan K Vester; Rolle Rahikainen; Irsyad N A Khairil Anuar; Rory A Hills; Tiong Kit Tan; Mark Howarth Journal: Nat Commun Date: 2022-06-28 Impact factor: 17.694
Authors: Craig Sweet; Aayush Aayush; Logan Readnour; Kevin V Solomon; David H Thompson Journal: Biomacromolecules Date: 2021-04-07 Impact factor: 6.988