Single-step purification of proteins of interest from proteolytically cleaved recombinant maltose-binding protein (MBP) fusion proteins by selective immunoprecipitation of MBP
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- Single-step purification of proteins of interest from proteolytically cleaved recombinant maltose-binding protein (MBP) fusion proteins by selective immunoprecipitation of MBP
- Jung Hyun Park; Shin Young Na; Dong Gun Lee; Byoung Don Han; Kil Lyong Kim
- Bibliographic Citation
- Biotechnology and Bioprocess Engineering, vol. 3, pp. 82-86
- Publication Year
- The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated by centrifugation, resulting in highly purified proteins in the supernatant.
- maltose binding protein (MBP)factor Xarecombinant proteinprotein purificationanti-MBP monoclonal antibody
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- 1. Journal Articles > Journal Articles
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