Development of an Ultrasensitive Monoclonal Antibody-Based Time-Resolved Fluorescence Immunochromatographic Assay for Mebendazole Detection

IF 6.2 1区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Journal of Agricultural and Food Chemistry Pub Date : 2025-10-10 DOI:10.1021/acs.jafc.5c07301

Zhan-Peng Chen, Jie Liu, Lin Luo, Bao-Zhu Jia, Bin Lu, Wenwen Wang, Peiqiang Shuai, Zhen-Lin Xu

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Abstract

Mebendazole (MBZ), a broad-spectrum benzimidazole anthelmintic widely applied in veterinary medicine and aquaculture, has raised increasing concerns about its potential residues in foods of animal origin. In this work, a high-affinity and specific monoclonal antibody (mAb) was generated using a newly designed hapten containing a carboxylic spacer at the N1 site of the benzimidazole ring. The obtained mAb exhibited an IC₅₀ of 0.13 ng/mL with negligible cross-reactivity against related compounds. By combining this mAb with time-resolved fluorescent microspheres, a time-resolved fluorescence immunochromatographic assay (TRFIA) was developed for rapid and quantitative MBZ detection. The TRFIA achieved a detection limit of 1.02 μg/kg in fortified mutton and carp samples, with recovery rates ranging from 83.5% to 114.5% and coefficients of variation below 10.8%. Results correlated strongly with LC–MS/MS (R² > 0.97). This TRFIA provides a highly sensitive, reliable, and cost-effective platform for on-site screening of MBZ residues in animal-derived foods.

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基于单克隆抗体的超灵敏时间分辨荧光免疫层析检测甲苯达唑方法的建立

苯并咪唑（MBZ）是一种广泛应用于兽药和水产养殖的广谱苯并咪唑类驱虫药，其在动物源性食品中的潜在残留引起了越来越多的关注。在这项工作中，利用新设计的在苯并咪唑环N1位点含有羧基间隔物的半抗原产生了一种高亲和力和特异性的单克隆抗体（mAb）。所得单抗的IC50为0.13 ng/mL，与相关化合物的交叉反应性可忽略不计。通过将该单抗与时间分辨荧光微球结合，开发了一种时间分辨荧光免疫层析法（TRFIA），用于快速定量检测MBZ。TRFIA对强化羊肉和鲤鱼样品的检出限为1.02 μg/kg，回收率在83.5% ~ 114.5%之间，变异系数在10.8%以下。结果与LC-MS /MS呈正相关（R2 > 0.97）。该TRFIA为动物源性食品中MBZ残留的现场筛查提供了一个高度敏感、可靠和经济的平台。

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来源期刊

Journal of Agricultural and Food Chemistry 农林科学-农业综合

CiteScore

9.90

自引率

8.20%

发文量

1375

审稿时长

2.3 months

期刊介绍： The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.