Maltose binding protein (MBP) fusion proteins with low or no affinity to amylose resins can be single-step purified using a novel anti-MBP monoclonal antibody

• Published in: Molecules and cells 1998-12, Vol.8 (6), p.709-716
• Main Authors: Park, J H, Choi, E A, Cho, E W, Hahm, K S, Kim, K L
• Format: Article
• Language: English
• Subjects: Amylose - chemistry; Animals; Anion Exchange Resins - chemistry; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; ATP-Binding Cassette Transporters; Blotting, Western; Carrier Proteins - immunology; Carrier Proteins - isolation & purification; Chromatography, Affinity; Escherichia coli Proteins; Maltose-Binding Proteins; Mice; Mice, Inbred BALB C; Monosaccharide Transport Proteins; Recombinant Fusion Proteins - isolation & purification; Sepharose - chemistry
• ISSN: 1016-8478
• DOI: 10.1016/S1016-8478(23)13487-X

The maltose binding protein (MBP) fusion protein expression system is a powerful tool to produce and isolate recombinant proteins in E. coli. Whereas the conventional isolation technique for MBP-fusion proteins takes advantage of the binding affinity of MBP to maltose, this method is limited insofar as the biological activity of MBP has to be fully conserved for a successful purification. In this study, a novel monoclonal antibody (mAb) specific for MBP, termed HAM-19, was generated and its application in the purification and detection of MBP-fusion proteins determined. Using anti-MBP immunoaffinity columns, even recombinant MBP fusion products with lowered or impaired binding affinity to maltose were purified in a single step procedure. In comparison to amylose resins, HAM-19 immunoaffinity columns showed a higher binding capacity and affinity to MBP-fusion proteins. Furthermore, the mAb HAM-19 also provides a technical improvement over polyclonal antisera for the detection and analysis of MBP-fusion proteins which are under use in various forms in the fields of molecular and cellular biology.