Corpuz, Angelica Revibes creator Loc Thai Nguyen Chairperson Suwussa Bamrungsap Co-Chairperson Anal, Anil Kumar Examination Committee Salin, Krishna R. Examination Committee Asian Development Bank-Japan Scholarship Program (ADB-JSP) Scholarship donor text series technical report th Pathum Thani, Thailand Asian Institute of Technology 2021 continuing eng 156 leaves : ill. The drawbacks of the conventional gluten detection techniques have put biosensors in the spotlight. They can be functionalized to become highly sensitive, selective, simple and cheap detection platforms for gluten-free food testing and verification. In this study, nanomaterial amplification and PDA-based epitope imprinting were applied for voltammetric detection of gliadin, a known pathogenic component of gluten. Critical synthesis and sensing conditions were optimized based on current response while voltammetric response, SEM, Raman spectroscopy and FTIR confirmed the modification. The electrochemical synthesis proved to be effective in depositing conductive RGO film and then imprinted polymeric film on bare SPCE. It was crucial to optimize the thickness of the RGO (10 CV scans) and polymeric (20 CV scans) films, template concentration (0.5 g/L), pre-polymerization time (30 mins), elution time (30 mins), rebinding pH (6.5) and time (10 mins) for a highly sensitive and selective interface. The sensor was able to detect native gliadins in buffered matrix in the linear range of 0.5-15ppm (LOD 0.69, LOQ 2.29) with high selectivity for PQQPFPQQ template (IF=8.68) and gliadin (IF=3.14) and can discriminate against OVA (Ý=0.15), FA (Ý=0.25) and casein (Ý=0.39). It is stable at room temperature for 2 weeks (7.52% RSD), reproducible (2.97% RSD) and reusable (1.64% RSD) once. Negligible matrix effects and reproducible measurements were found in rice flour matrix with higher recovery rates (101.96% and 99.19% for 1.15 and 10 ppm, respectively) than that of ELISA test kit II of Morinaga Institute (82.61% and 83.50%). This suggested the accessibility of epitope on surface of native gliadin and the feasibility of using an acidic template for PDA-based imprinting. The developed technique was able to surpass performance of more sophisticated EG-FET biosensor and a label-free immunoassay demonstrating its robustness. Evaluation of applicability to other toxic prolamins, hydrolyzed or heat-processed gluten sources and other food matrixes were recommended. Finally, certification of the spiking standard material to attest its actual concentration and completion of three data points for proper linear correlation with ELISA is proposed. A thesis submitted in partial fulfillment of the requirements for the degree of Master of Engineering in Food Engineering and Bioprocess Technology, School of Environment, Resources and Development Thesis (M. Eng.) - Asian Institute of Technology, 2021 Proteins Analysis Electrochemistry Asian Institute of Technology. http://203.159.5.9/ait-thesis/detail.php?q=B14882 http://203.159.5.9/ait-thesis/detail.php?q=B14882 211006 20260818090900.0