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 Pha 2023
Accepted
17 Phe 2024
First published
15 Kol 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

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements