Monospecific and ultrasensitive detection of ofloxacin: A computational chemistry-assisted hapten screening strategy and analysis of molecular recognition mechanism

Jiaxu Xiao; Liangni Qin; Dan Zhao; Niexie Huang; Wennuo Xu; Linwei Zhang; Xiaoming Pan; Shiyun Han; Mingyue Ding; Long Li; Tao Le; Dapeng Peng

doi:10.1016/j.jhazmat.2023.133221

Monospecific and ultrasensitive detection of ofloxacin: A computational chemistry-assisted hapten screening strategy and analysis of molecular recognition mechanism

J Hazard Mater. 2024 Mar 5:465:133221. doi: 10.1016/j.jhazmat.2023.133221. Epub 2023 Dec 12.

Authors

Jiaxu Xiao¹, Liangni Qin¹, Dan Zhao¹, Niexie Huang¹, Wennuo Xu¹, Linwei Zhang¹, Xiaoming Pan¹, Shiyun Han¹, Mingyue Ding¹, Long Li¹, Tao Le², Dapeng Peng³

Affiliations

¹ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China.
² College of Life Sciences, Chongqing Normal University, Chongqing 401331, China. Electronic address: letao@cqnu.edu.cn.
³ National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan 430070, China; Shenzhen Institute of Nutrition and Health, Huazhong Agricultural University, Shenzhen 518000, China; Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China; Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China. Electronic address: pengdapeng@mail.hzau.edu.cn.

PMID: 38103295
DOI: 10.1016/j.jhazmat.2023.133221

Abstract

Contamination in food and the environment with fluoroquinolones (FQs) has become a serious threat to the global ecological balance and public health safety. Ofloxacin (OFL) is one of the most widely utilized sterilization agents in FQs. In the process of monitoring OFL, broad-spectrum monoclonal antibodies (mAb) cannot meet the demand for monospecific detection. Here, a computational chemistry-assisted hapten screening strategy was proposed in this study. Differences in the properties of antigenic epitopes were precisely extracted through a comprehensive comparative study of 16 common FQs molecules and a monospecific and ultrasensitive mAb-3B4 for OFL was successfully prepared. The screened fleroxacin (FLE) hapten was applied in a heterologous competition strategy resulting in a 20-fold improvement in the half inhibitory concentration (IC₅₀) of mAb-3B4 to 0.0375 μg L^-1 and cross-reacted only with marbofloxacin (MAR) in regulated FQs. In addition, a single-chain variable fragment (scFv) for OFL was constructed for the first time with an IC₅₀ of 0.378 μg L^-1. Molecular recognition mechanism studies validated the reliability of this strategy and revealed the key amino acid sites responsible for OFL specificity and sensitivity. Finally, ic-ELISA and GICA were established for OFL in real samples. This work provides new ideas for the preparation of monospecific mAb and improves the monitoring system of FQs.

Keywords: Computational simulation; Hapten screening; Molecular mechanism; Monospecificity; Ofloxacin; Residue detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anti-Bacterial Agents / chemistry
Computational Chemistry*
Enzyme-Linked Immunosorbent Assay
Fluoroquinolones
Haptens
Ofloxacin*
Reproducibility of Results

Substances

Ofloxacin
Fluoroquinolones
Haptens
Anti-Bacterial Agents