This study deals with the possibility of using bioactive substances to reduce the allergenic potential of food. In the first phase of the research, the work aimed to verify the inhibitory effect of bioactive substances on crustacean protein (tropomyosin) and gluten (gliadin) in model samples. In the next phase of the research, the inhibitory effect of selected antioxidants was demonstrated on samples from the market network. The ELISA method showed a decrease in the immunoreactivity of the kits (Veratox, Biocheck) by an average of 36% in the model samples. Model samples containing gluten and shellfish showed lower immunoreactivity due to the addition of p-coumaric acid (33.2%), β-carotene (26.5%), caffeic acid (33.3%), quercetin (26.4%), gallic acid (46.2%), lycopene (32.4%), epicatechin (43.5%), chlorogenic acid (32.7%), and rutin (46.8%). This study further demonstrated an inhibitory effect in samples from the market network with gluten content (breadcrumbs, tarragon) by an average of 61.1% and in samples with crustacean content (shrimp and surimi sticks) by 15.7% after the addition of lycopene, epicatechin, and caffeic acid. The results confirmed the impact of bioactive substances on the immunoreactivity of ELISA tests and provide a foundation for developing hypoallergenic foods and enhancing allergen detection accuracy in food safety applications.