摘要
通过建立双抗体夹心ELISA法测定食物中大豆过敏原蛋白成分,为进出口食品中大豆过敏原成分检测和食物过敏性疾病的预防提供技术基础。提取大豆总蛋白,免疫小鼠制备抗大豆总蛋白的多克隆抗体,用该抗体做包被抗体,并用生物素标记该多抗作为捕捉抗体,从而建立双多抗体夹心ELISA法;检测该方法的灵敏度,同时检测10种食品中是否含有大豆过敏原蛋白成分。SDS-PAGE电泳结果显示大豆过敏原总蛋白在120、60、35、30、28、18ku处条带明显;免疫印迹显示,这些条带与制备的高效价抗大豆蛋白的多抗具有明显的阳性反应。检测食品中大豆过敏原蛋白成分双抗体夹心ELISA法最低检出限为～100ng/mL,标准曲线在100ng/mL～12.8μg/mL范围内线性良好;10种进口食品检测结果与食物过敏原标签标注内容相符。
To develop a new method with sandwich-antibody enzyme linked immunosorbent assay （ELISA） to detect soybean allergen protein trace in food products, which will provide a technical tool to prevent soybean allergy. The total soybean protein was extracted and further analyzed by SDS-PAGE. The Balb/e mice were immuned with the extracts of soybean proteins and the highly-titrated polyelonal IgG antibodies against them were isolated and coupled with biotin for sandwich ELISA kit. Ten food products were tested using this method. SDS-PAGE showed that there were main protein bands such as 120,60,35,30,28 and 18 ku. Our results indicated that the standard curve was linear with soybean allergen protein concentration from 100 ng/mL-12.8 ug/mL and the detection limit was -100 ng/mL of soybean allergen protein. Using these ELISA kits, we detected 10 foods and they were consistent with the food allergen labels from their manufactures.
出处
《食品研究与开发》
CAS
北大核心
2009年第5期105-109,共5页
Food Research and Development
基金
深圳非共识项目(2007)
深港创新圈计划项目(2007)
