双抗夹心酶联免疫吸附法检测荞麦过敏蛋白  

Double-antibody sandwich enzyme linked immunosorbent assay for the detection of buckwheat allergen protein

作　　者：张新瑀 崔晓东[1] 杨欢[1] 葛川[2] 王转花[1] 

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]山西省科学技术情报研究所,山西太原030001

出　　处：《食品工业科技》2015年第8期53-56,共4页Science and Technology of Food Industry

基　　金：国家自然基金项目(31171659);青年科学基金项目(31300653);山西省科技平台项目(2014091028)资助

摘　　要：提取制备荞麦过敏原蛋白（TBt）并免疫动物,分别制备鼠多克隆抗体和兔多克隆抗体,建立了双抗夹心酶联免疫吸附检测法（ELISA）用于检测食品中的荞麦过敏原。SDS-PAGE结果表明,纯化的TBt纯度达到98%以上,鼠抗血清效价为1∶6400。建立的双抗夹心ELISA法对荞麦过敏原蛋白的最低检测限为0.16μg/m L,线性范围为0.16~16μg/m L。并采用建立的双抗夹心ELISA法对市场出售的15种食品中荞麦过敏成分进行了检测。结果显示,该方法对绝大多数食品中的荞麦过敏原都有很好的特异性及灵敏性,说明该法可用于食物过敏原的诊断及食品中微量荞麦过敏成分的检测。In the present study,buckwheat allergenic protein（TBt） from tartary buckwheat seeds was extracted and purified,and specific antibodies against TBt were produced. A double-antibody sandwich enzyme linked immunosorbent assay（ ELISA） based on rabbit polyclonal antibody and mouse polyclonal antibody was established. This method could be used for the detection of buckwheat allergenic protein in food. SDS-PAGE experiments showed that the purified TBt reached a purity of more than 98%. The serum antibody titers from mouse was 1∶6400. The sandwich ELISA could detect buckwheat allergenic protein at levels as low as 0.16μg/m L,and the linear range was at 0.16~16μg/m L. Using the ELISA to detect 15 kinds of food commercially available in the market,it was found that the newly established ELISA had specificity and sensitivity toward different foods expect vinegar. It provided a method for the diagnosis of food allergenic and the detection of trace allergens in food.

关 键 词：荞麦 过敏蛋白 多克隆抗体 酶联免疫吸附法 

分 类 号：TS207.3[轻工技术与工程—食品科学]