Luo Lin, Kang Rui-Yao, He Zhen-Xi, Jia Bao-Zhu, Chen Zi-Jian, Deng Hao, Xu Zhen-Lin
Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China.
Heyuan Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Heyuan 517000, China.
Food Chem X. 2024 Feb 28;22:101255. doi: 10.1016/j.fochx.2024.101255. eCollection 2024 Jun 30.
In this study, three eugenol fragment-containing haptens were synthesized, and a monoclonal antibody (mAb) selective for five commonly-found eugenol compounds (EUGs, i.e., eugenol, isoeugenol, methyl eugenol, methyl isoeugenol, and acetyl isoeugenol) was obtained. Based on this mAb, a broad-spectrum indirect competitive ELISA for high-throughput detection of five EUGs was developed. The detection limits for eugenol, isoeugenol, methyl eugenol, methyl isoeugenol and acetyl isoeugenol in both tilapia and shrimp samples were 25.3/ 50.6 μg/kg, 0.075/0.15 μg/kg, 0.48/0.96 μg/kg, 0.16/0.32 μg/kg, and 18.16/36.32 μg/kg, respectively. The recoveries for five EUGs ranged from 80.4 to 114.0 % with a coefficient of variation less than 11.5 %. Moreover, homology modelling and molecular docking were conducted to elucidate the interactions mechanism of mAb-EUGs. The work provides a promising tool for high-throughput screening of EUGs in aquatic products, which can serve as a benchmark for designing haptens and developing immunoassays for other small molecules.
在本研究中，合成了三种含丁香酚片段的半抗原，并获得了一种对五种常见丁香酚化合物（即丁香酚、异丁香酚、甲基丁香酚、甲基异丁香酚和乙酰异丁香酚）具有选择性的单克隆抗体（mAb）。基于该mAb，开发了一种用于高通量检测五种丁香酚化合物的广谱间接竞争ELISA。罗非鱼和虾样品中丁香酚、异丁香酚、甲基丁香酚、甲基异丁香酚和乙酰异丁香酚的检测限分别为25.3/50.6μg/kg、0.075/0.15μg/kg、0.48/0.96μg/kg、0.16/0.32μg/kg和18.16/36.32μg/kg。五种丁香酚化合物的回收率在80.4%至114.0%之间，变异系数小于11.5%。此外，还进行了同源建模和分子对接，以阐明mAb与丁香酚化合物的相互作用机制。该工作为水产品中丁香酚化合物的高通量筛选提供了一种有前景的工具，可为设计半抗原和开发其他小分子免疫分析方法提供参考。
