STUDY ON THE ALLERGENECITY OF MAJOR SOYBEAN ALLERGEN Gly m Bd 26K

我们之前利用大豆过敏的特应性皮炎患者对大豆中的过敏原进行了阐明。Gly m Bd 68K、30K和26K是大豆的主要过敏原。目前还没有关于Gly m Bd 26K的报道。因此，在本研究中，我们试图提纯该变应原并阐明其性质。此外，还建立了Gly-m-Bd-26K双抗体夹心法，并用于检测各种豆制品中的过敏原含量。脱脂大豆经不同层析纯化得到接近均一的Gly m Bd 26K。纯化的变应原为ASN型糖蛋白，相对分子质量为26 kDa。该变应原的糖基化位点被确定为ASN-20。用菠萝酶和辣根过氧化物酶等其他植物糖蛋白和抗过氧化物酶糖基来检测大豆敏感患者血清中的部分IgE抗体是否能识别过敏原的糖基。为了测定大豆制品中Gly-m-Bd26K的含量，制备了两种单抗。利用这些抗体，建立了检测过敏原的夹心ELISA法。用本方法可测定0.5-10 ng的过敏原。用该方法测定了豆制品中甘氨酸、Bd、26K的含量。目前的过敏原在豆腐、汤鱼、腐竹和冬虫夏草中浓度较高，而在豆腐、味增、纳豆和添加了SPI的产品如肉丸中未检测到过敏原。

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.