Moriyama Tatsuya, Yano Erika, Kitta Kazumi, Kawamoto Shin-ichi, Kawamura Yukio, Todoriki Setsuko
Department of Applied Biological Chemistry, Graduate School of Agriculture, Kinki University.
Biosci Biotechnol Biochem. 2013;77(12):2371-7. doi: 10.1271/bbb.130487. Epub 2013 Dec 7.
The levels of food allergens in gamma-irradiated soybean (0, 2.5, 5, 7.5, 10, 20, and 30 kGy) were investigated by immunoblotting and ELISA, using allergen-specific antibodies and patient serum. After 3 months of storage, Coomassie brilliant blue (CBB) staining indicated similar total protein profiles among the treatments, but that some proteins were degraded by irradiation at high doses. Immunoblotting with specific antibodies for major soybean allergens (β-conglycinin, Gly m Bd 30 K, soybean trypsin inhibitor, and Gly m 4) resulted in apparent band profiles and intensities that were not significantly changed by irradiation. Competitive inhibition ELISA analyses suggested that there were no significant changes in the allergen contents, except for a decrease in the soybean trypsin inhibitor. The patient IgE binding allergenic protein patterns were not changed by irradiation up to 30 kGy. ELISA using patient serum also revealed that the IgE reactivity to the irradiated soybean extract did not increase from the level of the control, but that the reactivity to some patient serum IgE was significantly decreased by irradiation.
采用免疫印迹法和酶联免疫吸附测定法(ELISA),使用过敏原特异性抗体和患者血清,研究了经伽马射线辐照(0、2.5、5、7.5、10、20和30千戈瑞)的大豆中的食物过敏原水平。储存3个月后,考马斯亮蓝(CBB)染色显示各处理间总蛋白谱相似,但高剂量辐照会使一些蛋白质降解。用针对主要大豆过敏原(β-伴大豆球蛋白、Gly m Bd 30K、大豆胰蛋白酶抑制剂和Gly m 4)的特异性抗体进行免疫印迹分析,结果显示辐照并未使明显的条带图谱和强度发生显著变化。竞争性抑制ELISA分析表明,除大豆胰蛋白酶抑制剂含量有所降低外,过敏原含量没有显著变化。高达30千戈瑞的辐照并未改变患者IgE结合过敏原蛋白模式。使用患者血清进行的ELISA还显示,辐照大豆提取物的IgE反应性并未高于对照水平,但辐照会使某些患者血清IgE的反应性显著降低。