Preparation of high-affinity and high-specificity monoclonal antibody and development of ultra-sensitive immunoassay for the detection of 1-amino-hydantoin, the metabolite of nitrofurantoin

文献类型： 外文期刊

第一作者： Yang, Hongfei

作者： Yang, Hongfei;Qin, Liangni;Wen, Hao;Ding, Mingyue;Zhang, Linwei;Xiao, Jiaxu;Liang, Jixiang;Liu, Yiting;Yu, Ziyan;Peng, Dapeng;Yang, Hongfei;Qin, Liangni;Wen, Hao;Ding, Mingyue;Zhang, Linwei;Xiao, Jiaxu;Liang, Jixiang;Liu, Yiting;Yu, Ziyan;Peng, Dapeng;Peng, Dapeng;Peng, Dapeng

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关键词： Hapten design; Nitrofurantoin; 1-Aminohydantoin; icELISA; Derivatisation; Monoclonal antibody

期刊名称：FOOD CHEMISTRY （ 影响因子：9.8； 五年影响因子：9.7 ）

ISSN： 0308-8146

年卷期： 2024 年 456 卷

页码：

收录情况： SCI

摘要： 1-Aminohydantoin (AHD), the residual marker of nitrofurantoin, is usually detected after derivatisation using the derivatisation reagent 2-nitrobenzaldehyde. Avoiding the antibody recognition of the derivatisation reagent is essential for the accurate detection of AHD residues. In this paper, a novel hapten called hapten D was designed, and then, a monoclonal antibody that did not recognise 2-nitrobenzaldehyde was prepared based on this novel hapten. An ultra-sensitive indirect competitive enzyme linked-immunosorbent assay (icELISA) was established under optimal conditions. The 50% inhibition concentration and limit of detection of AHD were 0.056 and 0.0060 ng/mL, respectively, which improved the sensitivity by 9-37-fold compared with the previously reported icELISA methods. The average recovery rates were 88.1%-97.3%, and the coefficient of variation was <8.6%. The accuracy and reliability of the icELISA were verified using liquid chromatography-tandem mass spectrometry. These results demonstrated that the developed icELISA is a useful and reliable tool.

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