免疫磁性捕获ELISA法检测甘草中甘草蛋白的研究  

Immunomagnetic separation ELISA for identification of protein in Radix Glycyrrhizae

作　　者：曹宇[1] 曹学琳[1] 田旺[1] 杨文博[1] 白钢[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《中草药》2004年第10期1109-1112,共4页Chinese Traditional and Herbal Drugs

基　　金：天津市自然科学重点基金资助项目(003805311);教育部回国留学人员启动基金[(2000)479号]

摘　　要：目的 制备甘草蛋白免疫磁性微球,并建立快速、精确的免疫磁性捕获ELISA法检测甘草蛋白。方法 采用种子聚合法合成聚苯乙烯磁性微球,并以兔抗甘草蛋白IgG抗体致敏,制备特异性捕获甘草特征蛋白的免疫磁性微球。以生物素标记抗体为示踪抗体,结合辣根过氧化物酶标亲和素建立ELISA检测系统,用于甘草药材和含甘草中成药中甘草蛋白的分析。结果 利用该方法对甘草药材和中成药中甘草蛋白抗原检测,检测灵敏度达到10ng/mL。结论 免疫磁性捕获ELISA检测技术方便、快速、准确,为生药的品种鉴定及中成药的质量控制提供一种新方法。Object To provide a quick and precise immunomagnetic separation enzyme-linked immuosorbent assay (IMS-ELISA) for protein detecting of Radix Glycyrrhizae by immunomagnetic microspheres (IMMS). Methods Using dispersion polymerization, polystyrene magnetic microspheres as carriers were prepared. IMMS coating with rabbit anti-Radix Glycyrrhizae protein (RGP) IgG can specifically capture RGP. Taking biotinylated anti-RGP rabbit IgG as a tracer antibody, together with horseradish peroxidase (HRP) labeled avidin, an ELISA on IMMS was developed. Results The method was specific to RGP antigen and could identify 10 ng/mL RGP antigen. Conclusion IMS-ELISA shows a new practicality of the unequivocal identification of effective constituent for its speedness, convenience, and accuracy.

关 键 词：甘草 甘草蛋白 免疫磁性微球 免疫磁性捕获ELISA技术 

分 类 号：R286.02[医药卫生—中药学]