Investigation of the interactions between beta-lactams and a metallo-beta-lactamase from bacillus cereus using a monoclonal antibody.
A monoclonal antibody recognizing the active site of a beta-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 beta-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 beta-lactams was developed based on the inhibition of antibody binding of all the beta-lactams studied. The assay is applicable to detection of beta-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 beta-lactamases and could prove a useful tool in future drug development.
Chambers
S J
SJ
Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, United Kingdom.
Wyatt
G M
GM
Morgan
M R
MR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001409:Bacillus cereus / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D051379:Mice; D008807:Mice, Inbred BALB C; D001618:beta-Lactamases / Q000276:immunology / Q000378:metabolism*; D047090:beta-Lactams / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D047090:beta-Lactams; C409201:beta-lactamase IMP-1; D001618:beta-Lactamases