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Open AccessArticle

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

by Caixia Zhang 1,2,3,†, Qi Zhang 1,2,4,†, Xiaoqian Tang 1,2,4, Wen Zhang 1,2 and Peiwu Li 1,2,3,4,*
1
School of Life Sciences, Hubei University, Wuhan 430062, China
2
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China
3
Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China
4
Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China
*
Author to whom correspondence should be addressed.
Both C. Zhang and Q. Zhang rank as the first authors.
Toxins 2019, 11(5), 280; https://doi.org/10.3390/toxins11050280
Received: 22 April 2019 / Revised: 11 May 2019 / Accepted: 12 May 2019 / Published: 20 May 2019
(This article belongs to the Special Issue Fungal Infestations in Humans, Animals, Crops)
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 µg/mL, with a linear range of 0.027–0.653 µg/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. View Full-Text
Keywords: anti-idiotypic nanobody; surrogate; Ochratoxin A; enzyme-linked immunosorbent assay anti-idiotypic nanobody; surrogate; Ochratoxin A; enzyme-linked immunosorbent assay
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Zhang, C.; Zhang, Q.; Tang, X.; Zhang, W.; Li, P. Development of an Anti-Idiotypic VHH Antibody and Toxin-Free Enzyme Immunoassay for Ochratoxin A in Cereals. Toxins 2019, 11, 280.

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