Development of allergen specific RAST and assay method using culture cells

使用培养细胞开发过敏原特异性 RAST 和测定方法

• 批准号: 07556030
• 负责人: OGAWA Tadashi
• 金额: $ 3.84万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556030/
• 关键词: Soybean; Allergen; Hypoallergenic; Allergen-disk; Atopic dermatisis; Monoclonal antibody; RBL-2H3; Food allergy; アレルゲンデイスク; RAST

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.

34-大豆油体相关蛋白是大豆中主要的致敏蛋白。我们已经研究了降低大豆制品的致敏性。(1)在开发低过敏性豆制品之前，我们首先制备了用于放射性过敏原吸附试验的过敏原特异性纸片，并使用两种不同的单克隆抗体F5和H6建立了用于定量分析豆制品中过敏原的夹心ELISA方法。此外，为了估计由患者IgE识别的表位序列的位置，使用患者血清分析酶消化和CNBr降解的变应原片段。(2)用鼠抗Gly m Bd 30 K和鼠粘膜肥大细胞系RBL-2 H3的抗血清，建立了一种评价大豆变应原致敏性的微量测定方法。以Gly m Bd 30 k为抗原，亚克隆免疫BALB/c小鼠，制备抗Gly m Bd 30 k的抗血清。所开发的测定方法被证明是有用的模拟IgE介导的I型变态反应，并高度敏感的检测食物中的过敏原。

项目成果

H.Hosoyama, A.Obata, N.bando, H.Tsuji, and T.Ogawa: "Epitope analysis of soybean major allergen Gly m Bd 30K recognized y the mouse monoclonal antibody using overlapping peptides." Biosci.Biotech.Biochem.60. 1181-1182 (1996)

H.Hosoyama、A.Obata、N.bando、H.Tsuji 和 T.Okawa：“大豆主要过敏原 Gly m Bd 30K 的表位分析使用重叠肽识别小鼠单克隆抗体。”

R.Yamanishi, H.Tsuji, N.bando, I.Yoshimoto, and T.Ogawa: "Micro-assay method for evaluating the allergenicity of the major soybean allergen. Gly m Bd 30K.with mouse antiserum and RBL-2H3 cells" Biosci.Biotech.Boichem.61. 19-23 (1997)

R.Yamanishi、H.Tsuji、N.bando、I.Yoshimoto 和 T.Okawa：“用于评估主要大豆过敏原 Gly m Bd 30K 的过敏性的微量测定方法。使用小鼠抗血清和 RBL-2H3 细胞”

N.Bando: "Identification of the glycocilation site of a major soybean allergen Gly m Bd 30K." Biosci.Biotech.Biochem.60. 347-348 (1996)

N.Bando：“主要大豆过敏原 Gly m Bd 30K 糖纤化位点的鉴定。”

R.Yamanishi,et al.: "Reduction of the allergenicity of soybean by treatment with proteinases" J.Nutr.Sci.Vitaminol.42. 581-587 (1996)

R.Yamanishi 等人：“通过蛋白酶处理降低大豆的过敏性”J.Nutr.Sci.Vitaminol.42。

M.Hessing,et al.: "Comparison of human lgE-binding soya bean allergenic protein Gly m I with the antigenicity profiles of calf anti-soya protein sera." Food Agric.lmmunol.,. 8. 51-58 (1996)

M.Hessing 等人：“人 lgE 结合大豆过敏蛋白 Gly m I 与小牛抗大豆蛋白血清抗原性谱的比较。”