Generating Monoclonal Antibodies against Buprofezin and Developing Immunoassays for Its Residue Detection in Tea Samples

Abstract

The synthesis of a hapten and antigen for the preparation of a monoclonal antibody (mAb) for buprofezin is described. The recognition mechanism of hapten and buprofezin by monoclonal antibodies (mAb-19F2) is described. The effectiveness of the mAb-19F2 immunoassay technique was assessed, and the effective detection of buprofezin in tea samples was achieved through the establishment of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and colloidal gold immunochromatography assay (GICA). The mAb-19F2 subtype was IgG 1 , with an IC 50 of 1.8 ng/mL and a linear range (IC 20 − IC 80 ) of 0.6−5.4 μg/L, and had a cross-reaction rate of less than 0.18% with 29 other pesticides (neonicotinoids and insect growth regulators). The study identified π−π stacking interactions between hapten and TYR-61 at the mAb-19F2 site and alkyl/phosphate interactions with TRP-105 and ARG-103. The ic-ELISA had an IC 50 of 12.9 ng/mL in green tea and 5.65 ng/mL in black tea, with a recovery rate of 92.4%−101.0% and RSD of 2.1%−4.8%. The GICA had a limit of detection (LOD) was 500 ng/mL, with the complete disappearance of the test lines visible to the naked eye. The limit of quantitation (LOQ, IC 20 ) was determined to be 16.8 ng/mL. Additionally, the developed GICA showed no cross-reactivity with neonicotinoid pesticides. The recovery rate of tea spiked recovered samples was 83.6%−92.2%, with an RSD of 5.3%−12.6%, and the results were consistent with the LC/MS method. This study is important for the real-time detection of buprofezin residues to ensure food safety and human health.

Quantitative determination of prothioconazole in wheat grain, soybean, and pond water based on a polyclonal antibody

“…20,21 These methods are more suitable for precise laboratory analysis. Currently, due to their advantages such as the large number of samples or rapid on-site detection, sensitivity, and low cost, 22,23 immunoassays have become increasingly popular as tools for analyzing pesticides in agricultural and environmental samples. Enzyme-linked immunosorbent assay (ELISA), which has been shown to be as sensitive as conventional colorimetric immunoassays, can also be used for the quantitative determination of compounds.…”