Development of a gold nanoparticle enhanced enzyme linked immunosorbent assay based on monoclonal antibodies for the detection of fumonisin B-1, B-2, and B-3 in maize

In this paper, three hybridoma cell lines that secrete monoclonal antibodies against fumonisin B-1 (FB1), specifically antibody subtypes IgA (heavy-chain) and kappa (light-chain), were obtained by immunization and cell cloning procedures. And the surfaces of gold nanoparticles (AuNPs) were modified with mercaptoundecanoic acid (MUA) to obtain AuNPs-MUA, which was used as a carrier for horseradish peroxidase (HRP)-goat anti-mouse IgA to develop an enhanced indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for the detection of FB1. The 50% inhibitory concentration (IC50) and the limit of detection (LOD, 15% inhibitory concentration, IC15) of this method were 0.93 +/- 0.058 and 0.078 +/- 0.013 g L-1, respectively. The detection sensitivity was approximately 10 times higher than that of the traditional IC-ELISA (IC50 is 9.57 g L-1). The cross-reactivity rates of AuNP enhanced IC-ELISA with FB1, FB2, and FB3 were 100%, 86.9%, and 79.5%, respectively, and did not cross with other mycotoxins. The total amount of fumonisins (FB1, FB2, and FB3) in actual contaminated maize samples was determined by AuNP enhanced IC-ELISA, and the results were consistent with liquid chromatography-tandem mass spectrometry (LC-MS/MS) with R-2 = 0.9989. AuNP enhanced IC-ELISA showed good accuracy and stability, which made it one of the solutions to quantitatively detect the total content of fumonisins (FB1, FB2, and FB3) in maize.