Ultrasensitive monoclonal antibodies specific to thermal stable-soluble proteins of buckwheat

Abstract

Buckwheat is considered a severe food allergen, and its adulteration and mislabeling cause serious health risks. For protecting consumers suffering from buckwheat allergy, a high-sensitivity detection method is necessary to accurately identify intentional or unintentional adulteration of buckwheat in processed foods. The study revealed that buckwheat contains a significant amount of thermally stable-soluble proteins (TSSPs), which keep antigenicity even after heat treatment. Therefore, we used TSSPs to produce three monoclonal antibodies (MAbs) specific to buckwheat. A MAbs cocktail solution was subjected to enhance the sensitivity of an indirect enzyme-linked immunosorbent assay (iELISA), and the LOD was 1 ng/mL. The MAbs cocktail solution based-iELISA can successfully detect buckwheat adulterated in processed foods. The results suggested that the TSSPs in buckwheat can be used as suitable immunogens, and MAbs produced can be used as bioreceptor to develop immunoassays and biosensors for detecting buckwheat in food facilities and processed foods.

Keywords: Allergen molecules; Buckwheat allergen; ELISA; Monoclonal antibody; Thermal Stable-Soluble Protein (TSSP).