Electrochemical tracking of gluten in marketed foods by using a recombinant antibody fragment based-platform.

Abstract

The only treatment to effectively manage celiac disease is the avoidance of gluten containing foods. Therefore, and given its high prevalence, it is of utmost importance to have reliable and efficient methods for the detection of gluten to ensure the well-being and quality of life of celiacs. This work presents the development of an electrochemical immunoplatform exhibiting many practical advantages including simplicity, reduced cost and high sensitivity for the screening of gluten-containing products. The methodology exploited the unique features offered by a recombinant antibody fragment with high affinity towards gliadin together with the use of magnetic microcarriers (MμCs) as scaffolds for the implementation of an indirect competitive immunoassay. Using amperometric transduction on disposable electrodes and the horseradish peroxidase/hydrogen peroxide/hydroquinone system, a dynamic range between 7.3 and 1982 ng mL was obtained for gliadin standards, with a limit of detection of 1.4 ng mL. The developed immunoplatform was successfully employed for the analysis of a variety of processed foodstuffs, demonstrating the ability to discriminate between gluten-free and gluten-containing foods according to the legislated threshold (20 mg kg of gluten). The agreement with the results provided by the R5-based ELISA and qPCR methods confirmed the suitability of the bioplatform as a competitive tool in terms of assay time (results in just 60 min after gliadin extraction) sensitivity and applicability, even at the point of need.