Development of allergen specific RAST and assay method using culture cells

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34-kDa Oil-body-associated protein in soybean has been identified as a major allergenic protein in soybean. We have been studied on the reduction of allergenicity of soybean products.(1) Prior to the development of hypoallergeic soybean products, we prepared first the allergen specific paper disk for the radioallergosorbent test and established the Sandwich-ELISA method for a quantitative analysis of the allergen in soybean products using two distinct monoclonal antibodies, F5 and H6. Moreover, to estimate the location of an epitope sequences recognized by patient IgE,the enzyme-digested and CNBr-degraded allergen fragments were analyzed using patient's sera.(2) Amicro-assay method for evaluating the allergenicity of soybean allergen was developed by using the mouse antiserum against Gly m Bd 30K and RBL-2H3, a rat mucosal mast cell line. The anti-serum against Gly m Bd 30k was prepared by subctaneously immunized BALB/c mice with the allergen. The develped assay method is shown to be useful for simulating IgE mediated type I allergy and to be highly sensitive for detecting the allergen in foods.