Production and characterization of monoclonal and recombinant antibodies against antimicrobial sulfamethazine

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Title
Production and characterization of monoclonal and recombinant antibodies against antimicrobial sulfamethazine
Author(s)
Z Y Yang; W B Shim; Min-Gon Kim; K H Lee; K S Kim; K Y Kim; C H Kim; S D Ha; D H Chung
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 17, no. 4, pp. 571-578
Publication Year
2007
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 45 ng/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 IC50 value by IC-ELISA with scFv antibody was 4.8 ng/ml, compared with 1.6 ng/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.
Keyword
antimicrobialELISAmonoclonal antibodyscFv antibodysulfamethazine
ISSN
1017-7825
Publisher
South Korea
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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