Journal of Microbiology and Biotechnology
The Korean Society for Microbiology and Biotechnology publishes the Journal of Microbiology and Biotechnology.
Journal of Microbiology and Biotechnology
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2007 ; 17(4): 571~578

AuthorYang, Zheng-You, Won-Bo Shm, Min-Gon Kim, Kyu-Ho Lee, Keun-Sung Kim, Kwang-Yup Kim, Cheol-Ho Kim, Sang-Do Ha, Duck-Hwa Chung
TitleProduction and Characterization of Monoclonal and Recombinant Antibodies Against Antimicrobial Sulfamethazine
PublicationInfo J. Microbiol. Biotechnol.2007 ; 17(4): 571~578
AbstractA 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.
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KeywordsELISA, sulfamethazine, antimicrobial, monoclonal antibody, scFv antibody
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