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An immunoassay for ochratoxin A using tetramethylrhodamine-labeled ochratoxin A as a probe based on a binding-induced change in fluorescence intensity

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Abstract

Ochratoxin A (OTA) is a mycotoxin that can cause health risks to human/animal health. Contamination by OTA can occur in various foods and agricultural products, so sensitive and rapid detection of OTA is crucial. We describe a simple and sensitive fluorescence immunoassay for OTA using tetramethylrhodamine (TMR)-labeled OTA as a fluorescent probe. We conjugated tetramethylrhodamine to OTA through a covalent reaction, and obtained three TMR-OTA isomer probes after purification by high-performance liquid chromatography. All of the fluorescent probes showed high binding affinity to the anti-OTA antibody. Binding of the TMR-OTA probe to the antibody induced strong fluorescence of TMR-OTA due to the possible change in the local environment of TMR caused by affinity binding. In the presence of OTA, the OTA target competitively displaced the bound TMR-OTA probe from the antibody, causing a decrease in fluorescence. Measuring the change in fluorescence enabled rapid detection of OTA. This method was selective and allowed the detection of 1 nM OTA, showing potential for rapid OTA analysis in applications.

Graphical abstract: An immunoassay for ochratoxin A using tetramethylrhodamine-labeled ochratoxin A as a probe based on a binding-induced change in fluorescence intensity

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Publication details

The article was received on 22 Sep 2019, accepted on 19 Nov 2019 and first published on 03 Dec 2019


Article type: Paper
DOI: 10.1039/C9AN01879D
Analyst, 2020, Advance Article

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    An immunoassay for ochratoxin A using tetramethylrhodamine-labeled ochratoxin A as a probe based on a binding-induced change in fluorescence intensity

    Y. Li, N. Zhang, H. Wang and Q. Zhao, Analyst, 2020, Advance Article , DOI: 10.1039/C9AN01879D

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