作　　者： ; ; ; ; ;

机构地区： 深圳大学生命科学学院过敏反应与免疫学研究所

出　　处： 《食品科技》 2010年第2期241-245,共5页

摘　　要： 目的:建立双抗体夹心时间分辨荧光免疫法[Time-resolved Fluoroimmunoassay(TRFIA)]测定食物中花生过敏原蛋白成分,为进出口食品中花生过敏原成分检测和由花生导致的食物过敏性疾病的预防提供技术基础。方法:提取花生蛋白,免疫小鼠制备抗花生总蛋白的多克隆抗体,用该抗体包被酶标板,并用生物素标记该多抗作为捕捉抗体,Eu3+标记的链酶亲和素和以β-二酮体为主的增强液等试剂,建立双抗夹心TRFIA方法,检测该方法的灵敏度,同时用于13种食品中花生过敏原蛋白成分检测。结果:初步地研制出双抗体夹心TRFIA法检测食品中花生过敏原蛋白成分,具有特异性,和其最低检出限为0.1ng/mL,标准曲线在0.1～160ng/mL范围内线性良好;11种食品检测结果与食物过敏原标签标注内容相符,而2种标示不详的食品检测结果均呈现阳性。 Objective： To develop a new method with sandwich-antibody time-resolved fluoroimmunoassay （TRFIA） to detect peanut allergens protein trace in food products, which will provide a technical tool to prevent peanut allergy. Method： The peanut proteins were extracted and further analyzed by SDS-PAGE. The Balb/c mice were immuned with the extracts of peanut proteins and the highly-titrated polyclonal IgG antibodies against them were isolated and coupled with biotin for TRFIA kit. Thirteen food products were tested using this method. Result： We have successfully developed TRFIA kit for detection of the peanut allergens protein trace in food products. Our results indicated that the standard curve is linear with concentration of peanut allergens protein from 0.1 ng/mL to 160 ng/mL and the detection limit is 0.1 ng/mL of peanut allergens protein. We found that eleven foods were detected with the peanut allergens protein, which were consistent with the peanut allergens labels from their manufactures. However, two foods without clear food allergen label were detected to contain peanut allergens protein.

关 键 词： 时间分辨荧光免疫法 花生过敏原 多克隆抗体 食物过敏

领　　域： [轻工技术与工程] [轻工技术与工程]