Aminated PET Thin Film as a Functionalized Insulating Layer for Capacitive Gliadin Aptasensor Construction

Celiac patients require strict avoidance of gliadin, the primary immunotoxic component of gluten, making sensitive detection essential for food safety. A label-free and reagentless capacitive aptasensor for gliadin detection was developed using an aminated polyethylene terephthalate (PET) thin film as both an insulating layer and functionalization platform. The PET surface was modified via ethylenediamine-mediated aminolysis, enabling covalent immobilization of 5'-NH₂-modified gliadin aptamers through glutaraldehyde crosslinking. Under optimized conditions (23 µm initial PET thickness and 10 µM aptamer), the sensor showed a linear response from 10 to 500 µg/mL gliadin (R² = 0.9792), with a detection limit of 6.0 µg/mL, equivalent to 12 ppm gluten, which is well below the regulatory threshold of 20 ppm for gluten-free labeling. The aptasensor showed excellent correlation with commercial ELISA for 20 gluten-containing soy sauce samples (R² = 0.926) and spike recoveries of 91.7-105.7% in two gluten-free products. Efficient regeneration was achieved with 25 mM arginine (pH 9.0), retaining >80% activity after six cycles. This simple, low-cost, and reusable platform relies solely on a single PET thin film as consumable in a custom-built system with lab-friendly aminolysis conditions. It substantially lowers barriers to functionalized insulating layer fabrication, the primary challenge in capacitive aptasensor development, providing a promising method for on-site gliadin monitoring in gluten-free food safety applications.