Hapten synthesis, monoclonal antibody generation, and development of competitive immunoassays for the analysis of Picoxystrobin (cas 117428-22-5) in beer

Add time:07/20/2019    Source:sciencedirect.com

This paper describes the original synthesis of a functionalized derivative of the fungicide Picoxystrobin (cas 117428-22-5) and the generation of the first reported monoclonal antibodies against this strobilurin pesticide. The synthetic hapten was prepared by total synthesis from commercial chemicals and incorporating the spacer arm through a carbon–carbon single bond. Also, to obtain the immunogen, an uncommon hapten activation strategy based on N,N′-disuccinimidyl carbonate was employed, affording high activation yields and clean and reproducible coupling results. With these immunoreagents, two enzyme-linked immunosorbent assays (ELISAs) were developed: a competitive one-step assay using the antibody-coated direct ELISA format and a competitive two-step assay with the conjugate-coated indirect ELISA procedure. Both immunoassays were characterized in terms of sensitivity, selectivity, tolerance to solvents and matrix effects, achieving limits of detection below 0.2 μg L−1. The optimized assays were used for the determination of picoxystrobin residues in beer, with recovery values ranging between 90 and 121% for the direct assay and from 79 to 122% for the indirect assay.

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