Browse > Article
http://dx.doi.org/10.4014/jmb.1109.09053

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)
Publication Information
Journal of Microbiology and Biotechnology / v.22, no.2, 2012 , pp. 226-229 More about this Journal
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
Alexa-488; fusion; monoclonal antibody; protein chip; VHb;
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 Frey, A. D. and P. T. Kallio. 2003. Bacterial hemoglobins and flavohemoglobins: Versatile proteins and their impact on microbiology and biotechnology. FEMS Microbiol. Rev. 27: 525-545.   DOI   ScienceOn
2 Isarankura-Na-Ayudhya, C., S. Yainoy, T. Tantimongcolwat, L. Bulow, and V. Prachayasittikul. 2010. Engineering of a novel chimera of superoxide dismutase and Vitreoscilla hemoglobin for rapid detoxification of reactive oxygen species. J. Biosci. Bioeng. 110: 633-637.   DOI   ScienceOn
3 Jung, J. M., Y. B. Shin, M. G. Kim, H. S. Ro, H. T. Jung, and B. H. Chung. 2004. A fusion protein expression analysis using surface plasmon resonance imaging. Anal. Biochem. 330: 251-256.   DOI   ScienceOn
4 Khang, Y. H., I. W. Kim, Y. R. Hah, J. H. Hwangbo, and K. K. Kang. 2003. Fusion protein of Vitreoscilla hemoglobin with Damino acid oxidase enhances activity and stability of biocatalyst in the bioconversion process of cephalosporin C. Biotechnol. Bioeng. 82: 480-488.   DOI   ScienceOn
5 Kim, S. W., M. G. Kim, H. A. Jung, K. H. Lee, H. S. Lee, and H. S. Ro. 2008. An application of protein microarray in the screening of monoclonal antibodies against the oyster mushroom spherical virus. Anal. Biochem. 374: 313-317.   DOI   ScienceOn
6 Kwon, S. Y., Y. J. Choi, T. H. Kang, K. H. Lee, S. S. Cha, G. H. Kim, et al. 2005. Highly efficient protein expression and purification using bacterial hemoglobin fusion vector. Plasmid 53: 274-282.   DOI   ScienceOn
7 Park, K. W., D. A. Webster, B. C. Stark, A. J. Howard, and K. J. Kim. 2003. Fusion protein system designed to provide color to aid in the expression and purification of proteins in Escherichia coli. Plasmid 50: 169-175.   DOI   ScienceOn
8 Stark, B. C., K. L. Dikshit, and K. R. Pagilla. 2011. Recent advances in understanding the structure, function, and biotechnological usefulness of the hemoglobin from the bacterium Vitreoscilla. Biotechnol. Lett. 33: 1705-1714.   DOI   ScienceOn
9 Wakabayashi, S., H. Matsubara, and D. A. Webster. 1986. Primary sequence of a dimeric bacterial haemoglobin from Vitreoscilla. Nature 322: 481-483.   DOI   ScienceOn