Alkaline phosphatase triggered gold nanoclusters turn-on fluorescence immunoassay for detection of Ochratoxin A

Abstract

Ochratoxin A (OTA) is a highly toxic mycotoxin which can cause a variety of diseases. Sensitive detection of OTA is significant for food safety. Herein, a feasible and sensitive immunoassay was established for OTA detection by alkaline phosphatase (ALP) triggered gold nanoclusters (AuNCs) turn-on fluorescence. The fluorescence of the AuNCs can be quenched by Cr6+ induced aggregation of AuNCs and the fluorescence resonance energy transfer (FRET) between AuNCs and Cr6+. Under the catalytic action of ALP-labelled IgG (IgG-ALP), the ascorbic acid 2-phosphate (AA2P) was hydrolyzed to ascorbic acid (AA) for the reducing of Cr6+ to Cr3+. As a result, the degrees of AuNCs aggregation and FRET were weakened and the fluorescence of AuNCs was turned on. The amount of OTA in the sample was negatively correlated with the amount of IgG-ALP captured by anti-OTA monoclonal antibody (McAb) in the microplate. In optimal conditions, the turn-on fluorescence immunoassay had a good linear range of 6.25–100 ng/mL, and the detection limit was 0.693 ng/mL. The recoveries of OTA from corn were 95.89%-101.08% for the fluorescence immunoassay. This work provided a feasible, sensitive and good selectivity fluorescence method for OTA detection.