Abstract
In this work, a novel magnetic beads (MBs)-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2) is reported. It involves the use of sandwich-type immunoassays using selective capture and detector antibodies and carboxylic acid-modified magnetic beads (HOOC-MBs). Amperometric detection at −0.20 V was performed using dual screen-printed carbon electrodes (SPdCEs) and the H2O2/hydroquinone (HQ) system. This methodology exhibits high sensitivity and selectivity for the target proteins providing detection limits of 18.0 and 0.07 ng/mL for Ara h 1 and Ara h 2, respectively, with an assay time of only 2 h. The usefulness of the approach was evaluated by detecting the endogenous content of both allergenic proteins in different food extracts as well as trace amounts of peanut allergen (0.0001% or 1.0 mg/kg) in wheat flour spiked samples. The developed platform provides better Low detection limits (LODs) in shorter assay times than those claimed for the allergen specific commercial ELISA kits using the same immunoreagents and quantitative information on individual food allergen levels. Moreover, the flexibility of the methodology makes it readily translate to the detection of other food-allergens.
Highlights
Food allergies, i.e., adverse immunologic (IgE and non-IgE mediated) reactions to food, have resulted in considerable morbidity and reached high proportions in the industrialized world, affecting up to 10% of young children and 2%–3% of adults [1]
The magnetic beads (MBs) bearing the sandwich immunocomplexes for each target protein were magnetically captured on the corresponding working electrode (WE 1 and WE 2) of the screen-printed carbon electrodes (SPdCEs) and amperometric detection at0.20 V of the catalytic currents generated upon H2 O2 addition and using HQ as redox mediator in solution at each working electrode was employed to determine each target protein concentration
It is important to note that this methodology implied that the SPdCEs acted only as the electrochemical transducer while all the affinity reactions occurred on the surface of the MBs, minimizing unspecific adsorptions of the bioreagents on the electrode surfaces
Summary
I.e., adverse immunologic (IgE and non-IgE mediated) reactions to food, have resulted in considerable morbidity and reached high proportions in the industrialized world, affecting up to 10% of young children and 2%–3% of adults [1]. Analysis for food allergens is required both for consumer protection and food fraud identification. Peanut allergy deserves particular attention because very small amounts of peanut proteins can induce severe allergic reactions. It persists throughout life and accounts for most of food-induced anaphylactic reactions with a prevalence that has doubled in a five-year time span [3,4]. There is an increasing concern and need to protect food allergic consumers from acute and potentially life-threatening allergic reactions through detection of peanut trace contamination and accurate food labeling [5]. Regulation No 1169/2011 established food allergen labelling and information requirements under the EU Food Information for Chemosensors 2016, 4, 11; doi:10.3390/chemosensors4030011 www.mdpi.com/journal/chemosensors
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