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

主要大豆过敏原Gly m Bd 26K的过敏性研究

基本信息

批准号：
08660161
负责人：
TSUJI Hideaki
金额：
$ 1.47万
依托单位：
Okayama Prefectural University (1997)The University of Tokushima (1996)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660161/
关键词：
Sovbean Allergen Gly m Bd 26K Sugar chain soybec allergen cDNA cloning allergenicity

项目摘要

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.

我们之前曾利用大豆敏感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 被证明是主要的大豆过敏原。尚未报道有关 Gly m Bd 26K 的信息。因此，在本研究中，我们试图纯化过敏原并阐明其特性。此外，还开发了 Gly m Bd 26K 夹心 ELISA，并用于测定各种大豆产品中过敏原的含量。通过各种色谱法将脱脂大豆中的 Gly m Bd 26K 纯化至接近均质。纯化的过敏原是一种 Asn 型糖蛋白，分子量为 26 kDa。确定过敏原的糖基化位点为Asn-20。使用其他植物糖蛋白如菠萝蛋白酶和辣根过氧化物酶以及抗过氧化物酶的糖部分来检查大豆敏感患者血清中的部分IgE抗体是否能够识别过敏原的糖部分。血清中的部分IgE抗体可以与Gly m Bd 26K的糖部分结合。为了测定豆制品中Gly m bd26K的含量，制备了两种单克隆抗体。使用这些抗体，建立了针对过敏原的夹心 ELISA。通过本方法，可以测定0.5-10ng的过敏原。利用该方法测定了豆制品中Gly m Bd 26K的含量。目前的过敏原在豆腐、冬豆腐、腐竹和油豆腐中含量较高，但在酱油、味噌、纳豆以及添加了 SPI 的产品（如肉丸）中未检测到过敏原。