双抗体夹心ELISA法测定食物中虾过敏原成分

Detection of shrimp allergen protein trace in food products by sandwich-antibody enzyme linked immunosorbent assay

通过建立双抗体夹心ELISA法测定食物中虾过敏原成分,为食物过敏的预防与诊断提供理论基础。提取虾中过敏原蛋白,免疫小鼠制备抗虾过敏原的多克隆抗体,并以多克隆抗体包被酶标板,生物素标记多抗,建立双多抗体夹心的ELISA法测定食物中虾过敏原成分。成功地研制出双抗体夹心ELISA法检测食品中虾过敏原蛋白成分,具有特异性,其最低检出限为40ng/mL,标准曲线在40～1000ng/mL范围内线性良好。初步建立了检测虾过敏原蛋白成分的ELISA法,并具有较好的特异性和灵敏度,为进一步研制虾过敏原成分的ELISA检测试剂盒打下了基础。

Objective： To develop a new method with sandwich-antibody enzyme linked immunosorbent assay （ELISA）to detect shrimp allergen protein trace in food products, which will provide a technical tool to prevent shrimp allergy. Method： The total shrimp protein was extracted and further analyzed by SDS-PAGE. The Balb/c mice were immuned with the extracts of shrimp proteins and the highly-titrated polyclonal IgG antibodies against them were isolated and coupled with biotin for sandwich ELISA kit. Result： Our results indicated that the standard curve is linear with shrimp allergen protein concentration from 40-1000 ng/mL and the detection limit is 40 ng/mL of shrimp allergen protein. Thus we concluded that sandwich-antibody ELISA could be used to detect shimp allergen protein trace in food products.