Ultrasensitive antibody production strategy based on hapten property for simultaneous immunoassay

Wenjuan Wu; Changsheng Li; Desheng Liu; Jiawen Ji; Jianhui Zhu; Huizhe Lu; Bin Fu; Yongqiang Ma

doi:10.1016/j.foodchem.2022.133565

Ultrasensitive antibody production strategy based on hapten property for simultaneous immunoassay

Food Chem. 2022 Nov 30:395:133565. doi: 10.1016/j.foodchem.2022.133565. Epub 2022 Jun 23.

Authors

Wenjuan Wu¹, Changsheng Li¹, Desheng Liu¹, Jiawen Ji¹, Jianhui Zhu¹, Huizhe Lu¹, Bin Fu², Yongqiang Ma¹

Affiliations

¹ College of Science, China Agricultural University, Beijing 100193, China.
² College of Science, China Agricultural University, Beijing 100193, China. Electronic address: fubinchem@cau.edu.cn.

PMID: 35763926
DOI: 10.1016/j.foodchem.2022.133565

Abstract

A high-quality antibody production strategy is significant for immunoassay. In this work, four general haptens were proposed based on the 3D structure and surface electrostatic potential of molecular modeling. It was found that the sensitivity and specificity of polyclonal antibodies (pAbs) mainly depended on the bond angle of shapes liked "V" between haptens and proteins and hydrophobic parts of haptens. The quantified process was employed to obtain pAbs against cyhalofop-butyl and its metabolites (CAFs), with the IC₅₀ value of 4.9 μg·L^-1 under optimal conditions. The limit of quantization (LOQ) of the ultrasensitive icELISA in brown rice was 2 μg·kg^-1. The recoveries were 74%-110%, with a coefficient of variation (CV) less than 10%. This study indicated that the hapten property approach led to an improved immunoassay.

Keywords: Antibody production; Cyhalofop-butyl; Hapten; Molecular modeling; Simultaneous immunoassay.

MeSH terms

Antibodies
Antibody Formation*
Antibody Specificity
Enzyme-Linked Immunosorbent Assay
Haptens* / chemistry
Immunoassay
Models, Molecular

Substances

Antibodies
Haptens