Development of an Anti-Idiotypic VHH Antibody and Toxin-Free Enzyme Immunoassay for Ochratoxin A in Cereals

文献类型： 外文期刊

第一作者： Zhang, Caixia

作者： Zhang, Caixia;Zhang, Qi;Tang, Xiaoqian;Zhang, Wen;Li, Peiwu;Zhang, Caixia;Zhang, Qi;Tang, Xiaoqian;Zhang, Wen;Li, Peiwu;Zhang, Caixia;Li, Peiwu;Zhang, Qi;Tang, Xiaoqian;Li, Peiwu

作者机构：

关键词： anti-idiotypic nanobody; surrogate; Ochratoxin A; enzyme-linked immunosorbent assay

期刊名称：TOXINS （ 影响因子：4.546； 五年影响因子：4.8 ）

ISSN： 2072-6651

年卷期： 2019 年 11 卷 5 期

页码：

收录情况： SCI

摘要： Enzyme-linked immunosorbent assay (ELISA) test kits have been widely used for the determination of mycotoxins in agricultural products and foods, however, this test uses toxin standards with high toxicity and carcinogenicity that seriously threaten human health. In this work, the anti-idiotypic nanobody VHH 2-24 was first developed and then, using it as a surrogate standard, a toxin-free enzyme immunoassay for ochratoxin A (OTA) was established. The IC50 value of the VHH 2-24 surrogate standard-based ELISA was 0.097 mu g/mL, with a linear range of 0.027-0.653 mu g/mL. The average recoveries were tested by spike-and-recovery experiments, and ranged from 81.8% to 105.0%. The accuracy of the developed ELISA for detecting OTA was further verified by using the high performance liquid chromatography (HPLC) method, and an excellent correlation was observed. In summary, the toxin-free ELISA established in this study proves the latent use of the anti-idiotypic VHH as a surrogate calibrator for other mycotoxins and highly toxic small molecule analysis to improve assay properties for highly sensitive analyte determination in agricultural products.

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