Abstract
This work focused on the development and optimization of an impedimetric label-free immunosensor for detecting deoxynivalenol (DON). A monoclonal antibody for DON detection was immobilized on a modified gold electrode with a cysteamine layer and polyamidoamine (PAMAM) dendrimers. Cyclic voltammetry and electrochemical impedance spectroscopy techniques were used to monitor the layer-by-layer development of the immunosensor design, while electrochemical impedance spectroscopy and differential pulse voltammetry were employed to investigate the antigen/antibody interaction. The PAMAM dendrimers, allowing to immobilize a large number of monoclonal antibodies, permitted reaching, through the DPV technique, a high sensitivity and a low limit of detection equal to 1 ppb. The evaluation of the possible reuse of the immunosensors highlighted a decrease in the analytical performances of the regenerated immunosensors. After evaluating the matrix effect, the developed immunosensor was used to quantify DON in pasta samples spiked with a known mycotoxin concentration. Taking into consideration the DON extraction procedure used for the pasta samples and the matrix effect related to the sample, the proposed immunosensor showed a limit of detection of 50 ppb, which is lower than the maximum residual limit imposed by European Regulation for DON in dry pasta (750 ppb).
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