Broad-Specificity Immunoassays for Detecting Prochloraz and Its Metabolites in Grapes and Water


Zhao Y., Li P., HASSIBELNABY A. M. A., Lei X., Zhao J., Xu L., ...Daha Fazla

ACS Agricultural Science and Technology, 2025 (Scopus) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Basım Tarihi: 2025
  • Doi Numarası: 10.1021/acsagscitech.4c00648
  • Dergi Adı: ACS Agricultural Science and Technology
  • Derginin Tarandığı İndeksler: Scopus
  • Anahtar Kelimeler: broad-specificity antibody, computational chemistry analysis, immunoassay, prochloraz and its metabolites, recognition mechanism
  • Atatürk Üniversitesi Adresli: Evet

Özet

This study aimed to develop broad-specificity antibodies for detecting prochloraz and its metabolites. Initially, three haptens were designed and identified through computational chemical methods, and the hapten H2 was the optimal candidate. Using H2-BSA as an immunogen, the monoclonal antibody 9C3 was prepared to recognize PCH and two metabolites, BTS44595 and BTS44596. The broad-specificity recognition mechanism revealed that the “2,4,6-trichlorophenyl” moiety of the three targets is completely encapsulated in the binding pocket of mAb 9C3 and that hydrophobic forces play crucial roles in binding. Furthermore, an indirect enzyme-linked immunosorbent assay and a colloidal gold immunochromatographic assay for detecting prochloraz residues in grapes and river water samples were developed. The IC50 of ic-ELISA was 8.08-15.07 ng/mL, with recovery rates ranging from 73.80% to101.13%. In addition, the visual detection limit of GICA was 250-500 ng/mL, with IC50 values ranging from 98.04 to 126.29 ng/mL and CRs ranging from 67.2 to 78.02%. The accuracy and precision of both immunoassays were validated by LC-MS/MS.