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

文献类型： 外文期刊

第一作者： Xiao, Jiaxu

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

作者机构：

关键词： Ofloxacin; Computational simulation; Hapten screening; Monospecificity; Molecular mechanism; Residue detection

期刊名称：JOURNAL OF HAZARDOUS MATERIALS （ 影响因子：13.6； 五年影响因子：12.7 ）

ISSN： 0304-3894

年卷期： 2024 年 465 卷

页码：

收录情况： SCI

摘要： 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 mu g 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 mu g L-1. Molecular recognition mechanism studies validated the reliability of this strategy , 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.

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