Efficient purification of recombinant proteins fused to maltose-binding protein by mixed-mode chromatography.

Abstract

Two mixed-mode resins were evaluated as an alternative to conventional affinity resins for the purification of recombinant proteins fused to maltose-binding protein (MPB). We purified recombinant MBP, MBP-LacZ and MBP-Leap2 from crude Escherichia coli extracts. Mixed-mode resins allowed the efficient purification of MBP-fused proteins. Indeed, the quantity of purified proteins was significantly higher with mixed-mode resins, and their purity was equivalent to that obtained with affinity resins. By using purified MBP, MBP-LacZ and MBP-Leap2, the dynamic binding capacity of mixed-mode resins was 5-fold higher than that of affinity resins. Moreover, the recovery for the three proteins studied was in the 50-60% range for affinity resins, and in the 80-85% range for mixed-mode resins. Mixed-mode resins thus represent a powerful alternative to the classical amylose or dextrin resins for the purification of recombinant proteins fused to maltose-binding protein.

DOI: 10.1016/j.chroma.2009.03.048

Cite this paper

@article{Cabanne2009EfficientPO, title={Efficient purification of recombinant proteins fused to maltose-binding protein by mixed-mode chromatography.}, author={Charlotte Cabanne and J{\'e}r{\^o}me Pezzini and Gilles Joucla and Agn{\`e}s Hocquellet and Caroline Barbot and Bertrand Garbay and Xavier Santarelli}, journal={Journal of chromatography. A}, year={2009}, volume={1216 20}, pages={4451-6} }