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High-Efficiency Generation of Monoclonal Antibody for Vitreoscilla Hemoglobin Protein

  • Kim, Eun-Mi (Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Kim, Myung-Hee (Systems and Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Min-Gon (Graduate Program of Photonics and Applied Physics, GIST) ;
  • Kim, Sang-Woo (Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Ro, Hyeon-Su (Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University)
  • Received : 2011.09.21
  • Accepted : 2011.10.30
  • Published : 2012.02.28

Abstract

Bacterial hemoglobin from Vitreoscilla (VHb) is recognized as a good fusion protein for the soluble expression of foreign protein. In this study, we generated a monoclonal antibody (MAb) against VHb for its detection. For the rapid screening of MAb, a protein chip technology based on the Alexa-488 (A488) dye labeling method was introduced. In order to fabricate the chip, the VHb protein was chemically coupled to the chip surface and then the culture supernatants of 84 hybridoma cell lines were spotted onto the VHb chip. The bound MAbs were measured by A488-modified anti-mouse IgG. A single spot (MAb A10) exhibited significantly high signal intensity. The immunoblot analysis evidenced that the MAb A10 can detect VHb-fused proteins with high specificity.

Keywords

References

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