Preparation of monoclonal antibodies recognizing pharmacologically active metabolites of metamizole based on rational hapten design and their application in the detection of animal-derived food.

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2024-12-01 Epub Date: 2024-08-28 DOI:10.1016/j.talanta.2024.126753

Linwei Zhang, Jiacan Wang, Yiting Wang, Hao Wen, Mingyue Ding, Jiaxu Xiao, Hongfei Yang, Xiaoming Pan, Shiyun Han, Dapeng Peng

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Abstract

Metamizole (MET) is an antipyretic and analgesic drug, the illegal use of which can result in residues of MET metabolites in edible tissues of animals. In this study, a computational chemistry-assisted hapten screening strategy was used to screen for the optimal immunogenic hapten-A and the optimal coating antigen hapten-G-OVA. A monoclonal antibody capable of recognizing two pharmacologically active metabolites of MET, 4-methylamidinoantipyrine (MAA) and 4-aminoantipyrine (AA), was prepared from the hapten-A. The antibody showed excellent specificity for MAA and AA and almost no cross-reactivity with the pharmacologically inactive metabolites 4-formamidinoantipyrine (FAA) and 4-acetamidinoantipyrine (AAA). An ic-ELISA was developed for the simultaneous detection of MAA and AA in animal-derived food, the limits of detection for MAA ranged from 0.93 to 1.18 μg/kg, while those for AA ranged from 1.74 to 4.61 μg/kg. The recovery rate fell within the range of 82 %-110 %, with a coefficient of variation less than 16.39 %.

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基于合理的合体设计制备识别甲硝唑药理活性代谢物的单克隆抗体及其在动物源性食品检测中的应用。

甲硝唑（MET）是一种解热镇痛药，非法使用会导致动物可食用组织中残留甲硝唑代谢物。本研究采用计算化学辅助的单体筛选策略来筛选最佳免疫原性单体-A和最佳包被抗原单体-G-OVA。研究人员利用杂环素-A制备出了一种能够识别 MET 的两种药理活性代谢产物--4-甲脒基安替比林（MAA）和 4-氨基安替比林（AA）的单克隆抗体。该抗体对 MAA 和 AA 具有极好的特异性，与药理上无活性的代谢物 4-甲脒基安替比林（FAA）和 4-乙脒基安替比林（AAA）几乎没有交叉反应。该方法的检出限为 0.93 至 1.18 μg/kg，而 AA 的检出限为 1.74 至 4.61 μg/kg。回收率在 82 %-110 % 之间，变异系数小于 16.39 %。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.