Moiety and linker site heterologies for highly sensitive immunoanalysis of cyprodinil in fermented alcoholic drinks

Abstract

1Cyprodinil is a new-generation anilinopyrimidine fungicide widely used in crop protection 2 and frequently found in fruits. In this study, novel derivatives of cyprodinil with linker site 3 heterologies were synthesized and employed in order to produce antibodies with enhanced affinity. 4Moreover, moiety-heterologous haptens were designed and prepared for assay sensitivity 5 improvement. Two competitive enzyme-linked immunosorbent assays for the analysis of this active 6 substance were developed using direct and indirect formats, achieving IC 50 values around 0.15 7 µg/L. Analytical figures of merit and usability of the optimized assays were evaluated with wine and 8 cider as model food processed matrices. The obtained recoveries were from 90% to 120%, and the 9 limit of quantification was in the 1 to 5 µg/L range. Finally, a monitoring study (n=150) was 10

Monoclonal antibody-based immunoassays for cyprodinil residue analysis in QuEChERS-based fruit extracts

“…considering the good immune responses obtained previously in rabbits[Esteve-Turrillas et al, 2012;Esteve-Turrillas et al, 2015]. Moreover, these synthetic compounds hold the linker at opposite sites of the cyprodinil molecule, thus displaying different immunodeterminant chemical moieties at the distal end for antibody formation.…”

“…1). Previously, the immunogenicity of the different haptens was assessed by raising rabbit polyclonal antibodies (Esteve-Turrillas, Agulló, Abad-Fuentes, Abad-Somovilla, & Mercader, 2012;Esteve-Turrillas, Mercader, Agulló, Abad-Somovilla, & Abad-Fuentes, 2015). The aim of the present study was to generate high-affinity mAbs to cyprodinil and to develop and validate immunoassays for rapid and sensitive analysis of residues of this fungicide in QuEChERS-based fruit extracts.…”

Development a monoclonal antibody-based enzyme-linked immunosorbent assay for screening pyrimethanil in fruits and vegetables

“…The anilinopyrimidine class of fungicides are small molecules that must be conjugated to carrier proteins to render them immunogenic. Several haptens of anilinopyrimidines fungicide had been designed to obtain the special antibodies against pyrimethanil (Esteve-Turrillas et al, 2015b;Mercader et al, 2012) and/or cyprodinil (Esteve-Turrillas, Agullo, Abad-Fuentes, Esteve-Turrillas, Abad-Somovilla et al, 2015;Esteve-Turrillas et al, 2015a) in previous studies. In this study, the main aims of the hapten design were to produce the monoclonal antibody against pyrimethanil, and establish an ic-ELISA.…”

An impedimetric bioaffinity sensing chip integrated with the long-range DC-biased AC electrokinetic centripetal vortex produced in a high conductivity solution

“…Immunoassay relies on the use of specific antibodies to detect antigenic targets for various applications, 1,2 including clinical diagnostics, 3 food safety, 4 and the detection of environmental pollutants. 5 Label-free techniques, such as surface plasmon resonance, 6 quartz crystal microbalance, 7 mass-sensitive microcantilever, 8 and electrochemical impedance spectroscopy (EIS)based immunosensors, [9][10][11] have attracted increasing attention due to the reduced number of immunoassay steps, improved time efficiency, and reduced reagent cost.…”