From Computer to Sensor: A Proof-of-Concept Aptamer Platform Combining <i>In Silico</i> Design, Optical Validation, and Electrochemical Detection of the Fish Allergen <b>β</b> -Parvalbumin

Ensuring food safety is a critical concern in the modern food industry, as the presence of allergens, toxins, and pathogens can pose significant risks to consumer health. Beta-parvalbumin (β-PRVB) is a calcium-binding protein and the main fish allergen present in raw and processed food products, triggering severe allergic reactions in sensitive individuals. In this research paper, we explore the development of an electrochemical aptamer-based biosensor for the detection of β-PRVB. The biosensor development began with the <i>in silico</i> design of aptamers targeting β-PRVB, which were assessed for their docking and binding capabilities. This was followed by validation using a fluorescence quenching assay, ultimately leading to the creation of an electrochemical aptamer-based sensor utilizing the selected aptamer. The sensor showed a very low limit of detection (LOD) of 3.3 μg/mL and a limit of quantification (LOQ) of 10 μg/mL, with its specificity being tested with beta-lactoglobulin (β-Lact). Samples of commercial processed seafood products were analyzed to determine the effectiveness of the sensor, and the results were validated.