Production of monoclonal antibodies to the N-methylcarbamate pesticide propoxur.
Monoclonal antibodies (MAbs) to the pesticide propoxur were produced from haptens with carboxylic spacer arms of different lengths introduced at the carbamate group of the analyte structure. MAbs were subsequently characterized in the conjugate-coated format using these immunizing haptens and newly synthesized compounds as homologous and heterologous assay conjugates, respectively. Appropriate combinations of immunoreagents resulted in competitive enzyme immunoassays (ELISA) with I(50) values in the low nanomolar range (6.5-17.9 nM). A modification of the conjugate-coated format consisting of the simultaneous incubation of the MAb and the peroxidase-labeled secondary antibody in the presence of the analyte resulted in an assay with an I(50) value of 4.4 nM. This one-step conjugate-coated ELISA format is as simple and fast as the antibody-coated format but without the need of synthesizing enzyme-hapten conjugates. Major N-methylcarbamate pesticides were not recognized by the MAb. This immunoassay should reasonably allow the rapid, low-cost, and sensitive determination of propoxur in food, soils, and the environment at levels of regulatory and practical importance.
Moreno
M J
MJ
Laboratorio Integrado de Bioingeniería, Universidad Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain.
Abad
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Montoya
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000918:Antibody Specificity; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006241:Haptens / Q000276:immunology; D007114:Immunization; D007306:Insecticides / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D001074:Propoxur / Q000276:immunology*; D012710:Serum Albumin, Bovine / Q000276:immunology
D000911:Antibodies, Monoclonal; D006241:Haptens; D007306:Insecticides; D012710:Serum Albumin, Bovine; D001074:Propoxur