Production and Characterization of Monoclonal Antibodies Specific to the Strobilurin Pesticide Pyraclostrobin

Abstract

Strobilurin fungicides are nowadays among the most important fungicides in the market of active agrochemicals. Pyraclostrobin, which belongs to the last generation of this family of molecules, shows a broader antifungal activity spectrum and higher efficiency and security profiles than previous fungicides. This paper describes the synthesis of functionalized haptens, the production of monoclonal antibodies, and the development of enzyme-linked immunosorbent assays (ELISA) for the detection of pyraclostrobin. A conformational analysis of hapten structure was performed, which provided relevant data concerning the length of the spacer arm. A very useful strategy has been followed for the screening of hybridomas, leading to the selection of a panel of high-affinity monoclonal antibodies to pyraclostrobin. Moreover, different immunoassays have been characterized using the conjugate-coated indirect ELISA format, and limits of detection below 0.1 microg/L have been obtained. Also, a simplified one-step procedure has been carried out with two indirect assays. Finally, these results have been compared with the performance of the same antibodies in the antibody-coated direct ELISA format.

Hapten–antibody recognition studies in competitive immunoassay of α‐zearalanol analogs by computational chemistry and Pearson Correlation analysis

“…However, in most of the cases, antibody elicited by one hapten-carrier conjugate can cross-react with other molecules if these molecules share an identical or very similar epitope with the hapten used (Xu et al, 2006;Mercader et al, 2008). Actually, individual molecule always belongs to one class of compounds, for instance, sulfonamides family, thereby, the antibodies, particularly in the case of polyclonal antibodies, produced against one molecule often could recognize structurally similar analogs.…”

Synthesis and Fungicidal Activity of Novel Chloro-Containing 1-Aryl-3-oxypyrazoles with an Oximino Ester or Oximino Amide Moiety

“…Previous researchers have mainly focused on maintaining the pharmacophore but changing the side chain, and some new strobilurins such as trifloxystrobin, SYP-1620 and pyraclostrobin ( Figure 1) have been commercialized [5][6][7][8]. Our group has also devoted considerable effort to the development of novel fungicides.…”

Development of competitive enzyme-linked immunosorbent assays for boscalid determination in fruit juices

“…In order to find the proper immunoreagent concentration, a checkerboard competitive screening analysis for each antibody was performed in two immunoassay formats (d-and icELISA), as previously described (Mercader, Suárez-Pantaleón, Agulló, Abad-Somovilla, & Abad-Fuentes, 2008). For direct assays, plates were coated with several dilutions of the antiserum (from 1/10 4 to 1/10 5 ), and next day a range of tracer conjugate concentrations (from 3 to 100 ng/mL) was evaluated under competitive conditions.…”