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Combined heterologies for monoclonal antibody-based immunoanalysis of fluxapyroxad

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Abstract

Nowadays, instrumental methodologies and rapid bioanalytical techniques complement each other for the analysis of toxic chemical compounds. Fluxapyroxad was commercialized a few years ago as a fungicide and today it is being used worldwide to control a variety of pests. In the present study, the development of monoclonal antibody-based immunochemical methods for the analysis of this chemical in food samples was evaluated for the first time. Novel haptens were synthesized and protein bioconjugates were prepared. High-affinity and specific monoclonal antibodies to fluxapyroxad were generated from two haptens with alternative linker tethering sites. Haptens with linker site heterology and a structurally heterologous hapten with a minor modification of the molecule conformation and volume but with a significant alteration of the electronic density of the pyrazole moiety were evaluated for immunoassay development. Direct and indirect competitive immunoassays were characterized and optimized, showing IC50 values for fluxapyroxad of 0.14 and 0.05 ng mL−1, respectively. The combination of two heterologies was particularly adequate in the indirect format. The two developed immunoassays showed excellent recoveries and coefficients of variation in fluxapyroxad-fortified plums and four varieties of grapes. Finally, a good correlation was found between the indirect immunoassay and UPLC-MS/MS when fruit samples with incurred residues of fluxapyroxad were analyzed. These monoclonal antibody-based immunochemical methods hold great promise for fluxapyroxad monitoring.

Graphical abstract: Combined heterologies for monoclonal antibody-based immunoanalysis of fluxapyroxad

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Publication details

The article was received on 12 Sep 2018, accepted on 18 Oct 2018 and first published on 23 Oct 2018


Article type: Paper
DOI: 10.1039/C8AN01771A
Citation: Analyst, 2018, Advance Article
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    Combined heterologies for monoclonal antibody-based immunoanalysis of fluxapyroxad

    E. Ceballos-Alcantarilla, D. López-Puertollano, C. Agulló, A. Abad-Fuentes, A. Abad-Somovilla and J. V. Mercader, Analyst, 2018, Advance Article , DOI: 10.1039/C8AN01771A

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