A competitive immunoassay based on engineered magnetic/fluorescent nanoparticles and biolayer interferometry-based assay for T-2 toxin determination

For the first time a competitive immunoassay was developed by employing T-2 antibody-functionalized magnetite nanoparticles and T-2 toxin-conjugated fluorescent quantum dots (QDs). Free T-2 and the T-2-modified QDs compete for binding to antibody-modified magnetic beads; the magnetic beads collected by magnetic separation were subjected to fluorescence intensity analysis (with excitation/emission wavelengths at 460/616 nm). This competitive immunoassay for T-2 toxin determination was applied both in a microcentrifuge tube and on a 96-well plate. The dynamic range of the immunoassay is 1-100 ng mL(-1), the limit of detection (LOD) is 0.1 ng mL(-1), and determination was completed in about 40 min and 30 min in the microcentrifuge tube and 96-well plate, respectively. Moreover, the biolayer interferometry (BLI) technique was employed for T-2 determination for the first time, in which the conjugate of T-2 toxin and bovine serum albumin (BSA) was immobilized on the sensors before detection. Its average recovery of T-2 toxin from barley sample ranged from 82.00 to 123.33%, and the relative standard deviation (RSD) was between 9.42 and 15.73%. The LOD of the BLI-based assay is 5 ng mL(-1), and it only takes 10 min to finish the determination.