As a well-known allergenic indicator in kidney beans, lectins have always been the serious threats for human health. Herein, we introduced a new label-free voltammetric immunosensor for the direct determination of kidney bean lectin (KBL) with potential allergenic activity. Gold nanoparticles-polyethyleneimine-multiwalled carbon nanotubes nanocomposite was one-pot synthesized and modified onto the glass carbon electrode to enhance catalytic currents of oxygen reduction reaction. The KBL polyclonal antibody, acquired from rabbit immunization, was orientedly immobilized on the electrode modified with recombinant staphylococcal protein A via fragment crystallizable (Fc) region of antibody. Under the optimized condition, the immunosensor displayed a good linear response (R2 = 0.978) to KBL with a range from 0.05 to 100 μg/mL and a detection limit of 0.023 μg/mL. Simultaneously, the immunosensor exhibited well selectivity, interference-resistant ability, stability (4 °C) and reproducibility. Compared with the conventional enzyme-linked immunosorbent assay (ELISA) method, the immunosensor was successfully applied to quantify allergenic activity of lectin in raw and cooked (boiled for 30 min) kidney bean milk samples. This new approach provides new perspectives both for rapid quantification of lectin in kidney beans-derived foodstuffs and as a real-time monitoring tool for the allergenic potential during the whole production and consumption process.
Keywords: Allergenic activity detection; AuNPs-PEI-MWCNTs nanocomposite; Kidney bean lectin; Oriented antibody immobilization; Voltammetric immunosensor.
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