[关键词]
[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.
[Key word]
[Abstract]
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 (FIRM 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.
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[基金项目]
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]