NEB offers two bacterial protein expression systems in E.coli; the NEBExpress® MBP Fusion and Purification System (NEB #E8201) and the IMPACT™ System (NEB #E6901). Both systems can generate and purify high yields of recombinant proteins.
The NEBExpress® MBP Fusion and Purification System (NEB #E8201) takes advantage of the strong Ptac promoter and the translation initiation signals of maltose binding protein (MBP) to enhance solubility and expression levels of a desired protein in E. coli. The resulting product is an MBP fusion protein, which is then purified by affinity chromatography.
The IMPACT™ System (NEB #E6901) allows fusion of a tag consisting of an intein and a chitin binding domain (CBD) to either the C-terminus (pTXB1) or the N-terminus (pTYB21) of a target protein. In the presence of thiols, such as DTT, the intein undergoes specific self-cleavage, which releases the target protein from the chitin-bound intein tag resulting in purification of the target protein in a single chromatographic step with no need for proteases.
Over 40 years in protein expression and purification – a historical perspective
This article provides an overview of the advances in protein expression and purification methodology over the past 40 years.
- Protein Expression & Purification Brochure
- Purification Beads, Columns and Resins Brochure
- IMPACT™ Vectors and Applications
- Protein Expression and Purification Selection Chart
- Chong, S., Mersha, F.B., Comb, D.G., Scott, M.E., Landry, D., Vence, L.M., Perler, F.B., Benner, J., Kucera, R.B., Hirvonen, C.A., Pelletier, J.J., Paulus, H., and Xu, M.Q. (1997) Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element. Gene; 192, 271-281. PubMedID: 9224900
- Guan, C.D., Li, P., Riggs, P.D., and Inouye, H. (1988) Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein. Gene; 67, 21-30. PubMedID: 2843437
- Yu, H. H., et. al. (2010) Expressed protein ligation for the preparation of fusion proteins with cell penetrating peptides for endotoxin removal and intracellular delivery Biochim Biophys Acta; PubMedID: 20170629
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This webinar shares solutions developed by NEB scientists for producing various classes of protein, including: genetically-tailored E.coli host strains, expression vectors and valuable guidelines for generating properly-folded recombinant protein.