Broad-Specificity Immunoassays for Detecting Prochloraz and Its Metabolites in Grapes and Water
Zhao, Yun, Li, Peipei, Abd El-Aty, A. M., Lei, Xingmei, Zhao, Jing, Xu, Lingyuan, Gao, Song, Li, Jia, She, Yongxin, Jin, Fen, Wang, Jing, Zheng, Lufei, Hammock, Bruce D., Jin, Maojun
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.
- Dergi
- ACS AGRICULTURAL SCIENCE & TECHNOLOGY
- Yıl
- 2025
- Cilt
- 5
- Sayı
- 4
- Sayfa
- 528-541
- eISSN
- 2692-1952
- DOI
- 10.1021/acsagscitech.4c00648
- WoS UID
- WOS:001428478900001