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

Zhi-Li Xiao, Ya-Li Wang, Yu-Dong Shen, Zhen-Lin Xu, Jie-Xian Dong, Hong Wang, Chen Situ, Feng Wang, Jin-Yi Yang, Hong-Tao Lei, Yuan-Ming Sun

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In this paper, we report the development of a sensitiveand specific monoclonal antibody-based immunodiagnosticmethod for the detection of iso-tenuazonic acid (ITeA),an Alternaria mycotoxin, in food samples. The ITeA wasderivatized 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 glyoxalicacid 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 conjugationto bovine serum albumin (BSA). A highly specific monoclonalantibody selectively binding to ITeAH was generatedvia the hybridoma technique and subsequently used to constructa heterologous indirect competitive enzyme-linked immunosorbentassay (icELISA) using ITeAH as the competitiveantigen for the detection of ITeA with a limit of detection(LOD) of 0.5 ng/mL. Under the optimum conditions, the developedicELISA is highly sensitive (IC50 = 7.8 ng/mL) withrecovery rates ranged from 82.3 to 109.8% for spiked foodsamples. The comparative analysis of results revealed a goodcorrelation between the icELISA and the standard HPLC-MS/MS method, confirming the suitability of the developedicELISA for screening and detection of mycotoxin ITeA infood samples.
Original languageEnglish
Pages (from-to)635-645
Number of pages11
JournalFood Analytical Methods
Issue number3
Early online date14 Sep 2017
Publication statusEarly online date - 14 Sep 2017


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