Broad-Specificity Immunoassays for Detecting Prochloraz and Its Metabolites in Grapes and Water
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.
Authors
- Xingmei Lei
- Bruce D. Hammock (ORCID: https://orcid.org/0000-0003-1408-8317)
- Yongxin She (ORCID: https://orcid.org/0000-0001-7055-4808)
- Jing Zhao (ORCID: https://orcid.org/0000-0001-5177-5699)
- Maojun Jin (ORCID: https://orcid.org/0000-0002-9102-8572)
- Lingyuan Xu
- Fen Jin (ORCID: https://orcid.org/0000-0002-8686-7582)
- Yun Fu Zhao (ORCID: https://orcid.org/0000-0002-6357-9422)
- A.M. Abd El‐Aty (ORCID: https://orcid.org/0000-0001-6596-7907)
- Lufei Zheng
- Peipei Li (ORCID: https://orcid.org/0000-0003-0873-8475)
- Jing Wang (ORCID: https://orcid.org/0000-0002-8961-9277)
- Jia Li (ORCID: https://orcid.org/0000-0002-8610-5959)
- Song Gao (ORCID: https://orcid.org/0009-0005-8772-0733)
Institutions
- Cairo University (EG)
- University of California System (US)
- Jinhua Academy of Agricultural Sciences (CN)
- University of Veterinary Medicine (HU)
- Chinese Academy of Agricultural Sciences (CN)
- Ministry of Agriculture and Rural Affairs (CN)
- Institute of Quality Standards and Testing Technology for Agro Products (CN)
- UC Davis Comprehensive Cancer Center
- Atatürk University (TR)
- University of California, Davis (US)
Publication Details
- Journal
- ACS Agricultural Science & Technology
- Published
- 2025-02-21
- DOI
- https://doi.org/10.1021/acsagscitech.4c00648
- Primary Topic
- Pesticide Residue Analysis and Safety
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China
- Chinese Academy of Agricultural Sciences
- National Key Research and Development Program of China
- National Institute of Environmental Health Sciences