Issue 12, 2024

Heterologous antigen selection of chicken single-chain variable fragments against thiamethoxam

Abstract

Single-chain variable fragments (scFvs) are valuable in the development of immunoassays for pesticide detection. In this study, scFvs specific to thiamethoxam (Thi) were successfully isolated from a library generated by chicken immunization through heterologous coating selection. These scFvs were subsequently expressed with fusion with an Avi tag and alkaline phosphatase. After combination and optimization, a scFv-biotin based enzyme linked immunosorbent assay (ELISA) was developed for the detection of Thi, demonstrating an impressive half-maximum signal inhibition concentration (IC50) of 30 ng mL−1 and a limit of detection (LOD) of 1.8 ng mL−1. The immunoassay exhibited minimal cross-reactivity with other neonicotinoid insecticides, except for 7.5% for imidacloprid and 6.7% for imidaclothiz. The accuracy of the assay was confirmed by testing spiked samples of apple, pear, cabbage, and cucumber, which resulted in average recoveries ranging between 82% and 119%, closely aligning with the results obtained through high-performance liquid chromatography. Therefore, the chicken scFv-biotin based assay showed promise as a high-throughput screening tool for Thi in agricultural samples.

Graphical abstract: Heterologous antigen selection of chicken single-chain variable fragments against thiamethoxam

Supplementary files

Article information

Article type
Paper
Submitted
14 Leq 2023
Accepted
17 Qun 2024
First published
15 Nah 2024

Anal. Methods, 2024,16, 1756-1762

Heterologous antigen selection of chicken single-chain variable fragments against thiamethoxam

J. He, Y. Wang, D. Wang, Y. Yang, X. Xue, T. Xu, S. Gu and F. Tang, Anal. Methods, 2024, 16, 1756 DOI: 10.1039/D3AY01422C

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