Ovalbumin antibody-based fluorometric immunochromatographic lateral flow assay using CdSe/ZnS quantum dot beads as label for determination ofT-2 toxin.

Abstract

This work describes an anti-ovalbumin antibody-based lateral flow immunoassay (LFI) for T-2 toxin. The antibody uses acoating antigen as a bifunctional element for universality and introduces preincubation to improve the detection limits of the method. T-2 toxin and ovalbumin-modified T-2 toxin competitively binds on the anti-T-2 toxin monoclonal antibody modified on CdSe/ZnS quantum dot beads during preincubation. The modified T-2 toxin acts as a bifunctional element that forms immuno complexes during preincubation and combines with anti-ovalbumin antibody coated in the test line through the ovalbumin terminal. Fluorescence is detected at 610nm on the test zone following photoexcitation at 365nm. It has a reverse dose-effect relationship with the amount of T-2 toxin. The calibration plot is linear inthe 20-110fgmL-1T-2 toxin concentration range, and the limit of detection (LOD) is 10fgmL-1, which is lower by 8-fold than that of the traditional LFI system (LOD 80fgmL-1) and one order of magnitude than those of LFIs with labels of colloidal gold nanoparticles (LOD 150fgmL-1) or fluorophores (LOD 190ngmL-1). Universality was verified through aflatoxin B1 detection using the established ovalbumin antibody-based LFI system (LOD 10fgmL-1). The performance of the method was compared with that of established systems and a commercial ELISA kit (LOD 360fgmL-1). Graphical abstractSchematic representation of ovalbumin antibody-based immunochromatographic lateral flow assay for T-2 toxin. Preincubation is introduced for high sensitivity. T-2- anti-ovalbumin acts as a bi-functional element for universality. CdSe/ZnS quantum dot beads act as label. Fluorometric signal is detected at 610nm.