A broadly applicable approach to prepare monoclonal anti-cephalosporin antibodies for immunochemical residue determination in milk

Analytical and Bioanalytical Chemistry
Anna BremusErwin Märtlbauer

Abstract

A simple, efficient and rapid method for the synthesis of cephalosporin-protein conjugates was established. These conjugates were used as immunogens to produce monoclonal antibodies (mAbs) and as solid phase antigens in competitive indirect enzyme immunoassays (EIAs). With this generic approach, a novel set of monoclonal antibodies for cephalosporins was prepared, including ceftiofur and cephalexin as well as, reported here for the first time, cefoperazone, cefquinome and cephapirin. All 5 EIAs were highly sensitive, with standard curve IC(50) values of 0.7 (ceftiofur), 1.1 (cefquinome), 5.2 (cephalexin), 13.8 (cefoperazone) and 40.3 ng mL(-1) (cephapirin). Detection limits (IC(30)) ranged from 0.3 (ceftiofur mAb 1D7) to 17.2 ng mL(-1) (cephapirin mAb 2F10). Specificity studies revealed that cephalosporin-antibody binding was strongly determined by the side chain residues of the cephem nucleus. Therefore all mAbs, to some extent, recognized other beta-lactam antibiotics with similar side chain residues. Within the group of cephalosporins approved for use in veterinary medicine, however, the final EIAs were highly selective for their respective antigen, except for the ceftiofur EIA which showed cross-reactions with cefquinome. T...Continue Reading

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Citations

Sep 30, 2015·Analytical and Bioanalytical Chemistry·Natalia V Beloglazova, Sergei A Eremin
Sep 19, 2015·Mycotoxin Research·Richard Dietrich, Erwin Märtlbauer
Nov 9, 2017·American Journal of Industrial Medicine·Rolf MergetThomas Brüning
May 31, 2020·Bulletin of Environmental Contamination and Toxicology·Minxuan LiuXianghong Wang

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