Sandwich enzyme-linked immunosorbent assay system for micro-detection of the wheat allergen, Tri a Bd 17 K

Seven monoclonal antibodies (mAbs) against the wheat allergen, Tri a Bd 17 K, were prepared to obtain mAbs suitable for a sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for determination of the allergen. Two of the mAbs strongly immunoblotted the allergen purified from wheat flour. However, only one (1G11) of them was found to be suitable for sandwich ELISA. Epitope mapping against mAb-1G11 on the allergen showed that the mAb recognized the peptide containing Lys-38 and Gln-39 of the allergen. We developed a sandwich ELISA method consisting of Aleuria aurantia lectin for fixing the allergen and 1G11 as the first antibody that enabled 4-4,000 ng/well of the allergen to be determined.