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

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 nove...

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Bibliographic Details
Published in:Food chemistry 2024-10, Vol.456, p.140036, Article 140036
Main Authors: Yang, Hongfei, Qin, Liangni, Wen, Hao, Ding, Mingyue, Zhang, Linwei, Xiao, Jiaxu, Liang, Jixiang, Liu, Yiting, Yu, Ziyan, Peng, Dapeng
Format: Article
Language:English
Subjects:
1-Aminohydantoin
Derivatisation
Hapten design
icELISA
Monoclonal antibody
Nitrofurantoin
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Summary: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
ISSN:0308-8146
1873-7072
1873-7072
DOI:10.1016/j.foodchem.2024.140036