摘要
以β-伴大豆球蛋白免疫新西兰兔,制备抗β-伴大豆球蛋白多克隆抗体。利用正辛酸-硫酸铵沉淀法纯化多抗血清,并分析纯化前后多抗血清蛋白含量、纯度、效价、敏感性及特异性。结果表明,经聚丙烯酰胺凝胶电泳分析纯化后血清纯度得到提高;间接ELISA法检测多抗血清效价及敏感性无明显变化;与大豆Gly m Bd 28K蛋白和花生蛋白的交叉反应率分别由25.28%、1.16%降至为0.79%、0.2%。制得的抗β-伴大豆球蛋白多克隆抗体特异性高。为β-伴大豆球蛋白致敏蛋白的检测,和大豆β-伴大豆球蛋白致敏亚分子结构的定位奠定了基础。
Anti-β-conglycinin polyclonal antibody was generated in New Zealand rabbits using β-conglycinin as antigen and purifi ed by octanoic acid-ammonium surfate precipitation. And then serum protein content, purity, titer, sensitivity and specifi city before and after purifi cation were analyzed. The results showed that the serum protein content was increased by experiment of SDS-PAGE. The titer and sensitivity had no obvious change detected by indirect ELISA.The ratios of cross-reactivity of β-conglycinin with Gly m Bd 28 K and peanut protein were declined from 25.28% to 0.79% and 1.16% to 0.2%, respectively. The anti-β-conglycinin polyclonal antibody possessed high specifi city. The basis for the detection of β-conglycinin allergen and the identifi cation of epitopes were provided.
作者
皮江一
席俊
贺梦雪
闫慧丽
PI Jiang-yi;XI Jun;HE Meng-xue;YAN Hui-li(College of Food Science and Technology, Henan University of Technology, Zhengzhou 450001, Henan, China)
出处
《粮食与油脂》
北大核心
2018年第4期51-53,共3页
Cereals & Oils
基金
国家自然科学基金项目(31671778
31301409)
河南省高等学校重点科研项目(16A550001)
