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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.

我们以前进行了大豆过敏原的说明使用大豆过敏性患者特应性皮炎。Gly m Bd 68 K、30 K和26 K是大豆的主要过敏原。没有关于Gly m Bd 26 K的报告。因此，在本研究中，我们试图纯化过敏原，并阐明其性质。此外，还建立了Gly m Bd 26 K的双抗体夹心ELISA法，并用于各种豆制品中过敏原含量的测定。Gly m Bd 26 K是从脱脂大豆中通过各种色谱法纯化到接近均一的。纯化的过敏原被证明是一个天冬酰胺型糖蛋白，分子量为26 kDa。过敏原的糖基化位点被确定为Asn-20。用其他植物糖蛋白如菠萝蛋白酶、辣根过氧化物酶和抗过氧化物酶糖基抗体检测大豆过敏患者血清中IgE抗体的部分是否能识别过敏原的糖基。血清中的某些IgE抗体可与Gly m Bd 26 K的糖基部分结合。为了测定大豆制品中Gly m Bd 26 K的含量，制备了两株单克隆抗体。使用的抗体，夹心ELISA的过敏原已经建立。本方法可测定0.5- 10 ng的过敏原。用该法测定了大豆制品中Gly m Bd 26 K的含量。目前的过敏原被证明出现在高浓度的豆腐，tonyu，腐竹和abura年龄，但没有过敏原可以检测到酱油，味噌，纳豆和产品补充SPI，如肉丸。