Increasing the Solubility and Recovery of Ara h3 Allergen from Raw and Roasted Peanut

Abstract

Ara h3 belongs to the glycinin family of seed storage proteins and is one of the major peanut allergens. It comprises over 20% of the total peanut protein mass, making it a logical target for the detection of trace quantities of undeclared peanut contamination in foods. Both Ara h1 and Ara h3 are detected in lower quantities in cooked foods, either because of the failure to completely resolubilize the denatured proteins or because of the disruption of conformational epitopes required for monoclonal antibody recognition. A new reagent containing a proprietary non-detergent sulfobetaine (NDSB) is described which solubilizes more total protein and yields more Ara h3 protein from both raw and roasted peanut than other commonly used ELISA-compatible reagents.