Electrochemically assisted DNA and thioaromatic assembly as sensing and antifouling interface for food allergens

Food allergies have become a global issue and are estimated to affect approximately 220 million people worldwide. Allergy to peanuts can easily become life-threatening and induce anaphylactic reactions. Mislabeling and cross-contamination during food processing can occur in the frame of world population growth and pose a serious health issue. As the mandatory allergen list is not uniform worldwide, the development of routine analytical strategies with high specificity and sensitivity is a demanding task to aid in the rapid identification of allergenic foods. In this work, an electrochemical aptasensor for Ara h1 peanut allergen was developed by immobilizing the specific aptamer by the inserting method. First, a layer of p-aminothiophenol (p-ATP) was immobilized on the gold surface of screen-printed electrodes (GSPE) to improve the aptamer insertion and reduce the fouling effects at the electrode surface. The grafting of the p-ATP and Ara h1 aptamer on the GSPE surface was monitored by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). The resulting disposable aptasensor allowed for indirect electrochemical detection of Ara h1 protein in the presence of 5 mM ferro/ferricyanide as a redox probe. The electrochemical response upon aptamer-target interaction was monitored in the concentration range 1-250 nM, and two limits of detection in the nanomolar range were estimated based on DPV (2.78 nM Ara h1) and EIS (0.82 nM Ara h1) measurements. The aptasensor was successfully applied to real sample analysis.