Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production and Characterization of Monoclonal and Recombinant Antibodies Against Antimicrobial Sulfamethazine

  • Yang, Zheng-You (Division of Applied Life Science, Graduate School of Gyeongsang National University) ;
  • Shim, Won-Bo (Division of Applied Life Science, Graduate School of Gyeongsang National University) ;
  • Kim, Min-Gon (Laboratory of Integrative Biotechnology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Kyu-Ho (Department of Environmental Engineering and Biotechnology, Hankuk University of Foreign Studies) ;
  • Kim, Keun-Sung (Department of Food Science and Technology, Chung-Ang University) ;
  • Kim, Kwang-Yup (Department of Food Science and Technology, Chungbuk University) ;
  • Kim, Cheol-Ho (Department of Biochemistry and Molecular Biology, College of Oriental Medicine, Dongguk University and NRL-Glycobiology) ;
  • Ha, Sang-Do (Department of Food Science and Technology, Chung-Ang University) ;
  • Chung, Duck-Hwa (Division of Applied Life Science, Graduate School of Gyeongsang National University)
  • Published : 2007.04.30
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Abstract

A monoclonal antibody (mab) against the antimicrobial sulfamethazine was prepared and characterized by an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA). Sulfamethazine in the range of 0.2 and 45ng/ml could be determined with the mab by IC-ELISA. cDNAs encoding a variable heavy chain and variable light chain of the mab were cloned to produce recombinant antibodies using phage display technology. Following phage rescue and three rounds of panning, a single-chain variable fragment (scFv) antibody with high sulfamethazine-binding affinity was obtained. ELISA analysis revealed that scFv antibody and parent mab showed similar, but not identical, characteristics. The $IC_{50}$ value by IC-ELISA with scFv antibody was 4.8ng/ml, compared with 1.6ng/ml with the parent mab. Performances of the assays in the presence of milk matrix were compared; the mab-based assay was less affected than the scFv-based assay. Sixty milk samples were analyzed by mab-based IC-ELISA, and four samples were sulfamethazine positive; these results were favorably correlated with those obtained by HPLC.

Keywords

References

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