Double-antibody sandwich enzyme linked immunosorbent assay for the detection of buckwheat allergen protein

In the present study,buckwheat allergenic protein(TBt) from tartary buckwheat seeds was extracted and purified,and specific antibodies against TBt were produced. A double-antibody sandwich enzyme linked immunosorbent assay( ELISA) based on rabbit polyclonal antibody and mouse polyclonal antibody was established. This method could be used for the detection of buckwheat allergenic protein in food. SDS-PAGE experiments showed that the purified TBt reached a purity of more than 98%. The serum antibody titers from mouse was 1∶6400. The sandwich ELISA could detect buckwheat allergenic protein at levels as low as 0.16μg/m L,and the linear range was at 0.16~16μg/m L. Using the ELISA to detect 15 kinds of food commercially available in the market,it was found that the newly established ELISA had specificity and sensitivity toward different foods expect vinegar. It provided a method for the diagnosis of food allergenic and the detection of trace allergens in food.