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Specific Monoclonal Antibody-Based Enzyme Immunoassay for Sensitive and Reliable Detection of Alternaria Mycotoxin Iso-Tenuazonic Acid in Food Products

Xiao, Zhi-Li, Wang, Ya-Li, Shen, Yu-Dong, Xu, Zhen-Lin, Dong, Jie-Xian, Wang, Hong, Situ, Chen, Wang, Feng, Yang, Jin-Yi, Lei, Hong-Tao, Sun, Yuan-Ming
Food analytical methods 2018 v.11 no.3 pp. 635-645
Alternaria, analytical methods, bovine serum albumin, detection limit, enzyme-linked immunosorbent assay, foods, haptens, hybridomas, hydrazine, inhibitory concentration 50, monoclonal antibodies, mycotoxins, screening, tenuazonic acid
In this paper, we report the development of a sensitive and specific monoclonal antibody-based immunodiagnostic method for the detection of iso-tenuazonic acid (ITeA), an Alternaria mycotoxin, in food samples. The ITeA was derivatized with hydrazine hydrate to produce the antigen (E)-3-(1-hydrazonoethyl)-4-hydroxy-5-isobutyl-1H-pyrrol-2(5H)-one (ITeAH) which was further reacted with glyoxalic acid to generate the hapten (E)-2-((Z)-(1-(4-hydroxy-5-isobutyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl)ethylidene) (ITeAHGA) which was used as an immunogen after conjugation to bovine serum albumin (BSA). A highly specific monoclonal antibody selectively binding to ITeAH was generated via the hybridoma technique and subsequently used to construct a heterologous indirect competitive enzyme-linked immunosorbent assay (icELISA) using ITeAH as the competitive antigen for the detection of ITeA with a limit of detection (LOD) of 0.5 ng/mL. Under the optimum conditions, the developed icELISA is highly sensitive (IC₅₀ = 7.8 ng/mL) with recovery rates ranged from 82.3 to 109.8% for spiked food samples. The comparative analysis of results revealed a good correlation between the icELISA and the standard HPLC-MS/MS method, confirming the suitability of the developed icELISA for screening and detection of mycotoxin ITeA in food samples.