| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
We had previously carried out the elucidation of the allergens in soybean using soybean-sensitive patients with atopic dermatitis. Gly m Bd 68K,30K and 26K was shown to be the major soybean allergens. No information as to Gly m Bd 26K has been reported. Therefore, in the present study, we attempted to purify the allergen and to elucidate its properties. Furthermore, a sandwich ELISA for Gly m Bd 26K has been developed and utilized for the determination of the contents of the allergen in various soybean products. Gly m Bd 26K was purified to near homogeneity from defatted soybean by various chromatographies. The purified allergen was shown to be an Asn-type glycoprotein with a molecular mass of 26 kDa. The glycosylation site of the allergen was determined to be Asn-20. Whether some part of IgE antidbo dies in the sera of the soybean-sensitive patients can recognize the sugar moiety of the allergen was examined using other plant glycoproteins such as bromelain and horseradish peroxidase and anti-the sugar moiety of the peroxidase. Some IgE antibodies in the sera could bind to the sugar moiety of Gly m Bd 26K.
In order to determine the contents of Gly m bd26K in soybean products, two monoclonal antibodies have been prepared. Using the antibodies, a sandwich ELISA for the allergen has been established. By the present method, 0.5-10ng of the allergen could be determined. Using the method, the Gly m Bd 26K contents of soybean products were assayd. The present allergen was shown to occur at high concentrations in tofu, tonyu, yuba and abura-age, but no allergen could be detected in shoyu, miso, natto and products supplemented with SPI,such as meat balls.
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