数字农科院2.0

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

文献类型： 外文期刊

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

作者机构：

关键词： （0-1-3）1-Aminohydantoin;Derivatisation;Hapten design;icELISA;Monoclonal antibody;Nitrofurantoin

期刊名称： Food Chemistry

ISSN： 0308-8146

年卷期： 2024 年 456 卷

页码：

收录情况： SCIE（2024版） ; ; EI（2024版）

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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