Investigation of the Interactions between β-Lactams and a Metallo-β-lactamase from Bacillus cereus Using a Monoclonal Antibody

• Published in: Analytical biochemistry 2001-01, Vol.288 (2), p.149-155
• Main Authors: Chambers, S.J., Wyatt, G.M., Morgan, M.R.A.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Bacillus cereus - enzymology; beta-Lactamases - immunology; beta-Lactamases - metabolism; beta-Lactams - immunology; beta-Lactams - metabolism; Chromatography, Ion Exchange; Mice; Mice, Inbred BALB C
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1006/abio.2000.4883

A monoclonal antibody recognizing the active site of a β-lactamase from Bacillus cereus was identified and characterized. The binding of the monoclonal antibody to the active site was quantitatively inhibited by a broad spectrum of β-lactam antibiotics. The levels of inhibition were found to be associated with particular structural features of the antibiotics and their ability to form stable enzyme/substrate complexes. A novel, broad specificity assay for β-lactams was developed based on the inhibition of antibody binding of all the β-lactams studied. The assay is applicable to detection of β-lactams at or close to the MRL level and would be complementary to existing receptor-based assays. The approach described is relevant to the study of kinetic aspects of β-lactamases and could prove a useful tool in future drug development.