Development of a specific anti-idiotypic nanobody for monitoring aflatoxin M-1 in milk and dairy products

文献类型： 外文期刊

第一作者： Cai, Chong

作者： Cai, Chong;Zhang, Qi;Nidiaye, Seyni;Yan, Honglin;Zhang, Wen;Tang, Xiaoqian;Li, Peiwu;Cai, Chong;Zhang, Qi;Yan, Honglin;Tang, Xiaoqian;Li, Peiwu;Cai, Chong;Zhang, Qi;Yan, Honglin;Tang, Xiaoqian;Li, Peiwu;Cai, Chong;Zhang, Qi;Nidiaye, Seyni;Tang, Xiaoqian;Li, Peiwu;Cai, Chong;Zhang, Qi;Tang, Xiaoqian;Li, Peiwu

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关键词： Nanobody; Aflatoxin M-1; Phage-displayed; ELISA; Milk products

期刊名称：MICROCHEMICAL JOURNAL （ 影响因子：4.821； 五年影响因子：4.364 ）

ISSN： 0026-265X

年卷期： 2021 年 167 卷

页码：

收录情况： SCI

摘要： Anti-idiotypic antibodies (AIds) display stable protein conformation, offering an alternative to conventional antigens in immunoassays. In this work, an anti-idiotypic variable domain of heavy-chain antibody (VHH) or nanobody named VHH C4 binding to monoclonal antibody (mAb) 2C9 against aflatoxin M-1 (AFM(1)) was screened out from an alpaca immunized phage-displayed library. VHH C4 shows outstanding features in specificity and sensitivity, the IC50 is 0.25 ng/mL and exhibit almost no cross-reactivity with other aflatoxin analogs. As our knowledge, it is the first report for developing an anti-idiotypic nanobody for AFM(1). An environment-friendly competitive ELISA was developed using VHH C4 as a coating antigen. The results revealed a linear range from 0.10 ng/mL to 0.60 ng/mL, with a limit of detection (LOD) of 0.05 ng/mL. The recovery of spiked samples (pure milk, yogurt, and milk powder) ranged from 84.1% to 119.1%. The ELISA results of AFM(1) content in naturally contaminated samples were in good agreement with those determined by high performance liquid chromatography (HPLC). The results demonstrated that the nanobody VHH C4 is a promising alternative antigen in developing immunological methods for the detection of AFM1 in dairy products.

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