珍稀药源植物蛇足石杉中石杉碱甲的免疫学快速检测  

Establishment of Immunology Rapid Test Method for Huperzine A in Rare Source Plant Huperzia Serrata

作　　者：余宇燕[1] 张淑玲[1] 邹艳辉[1] 滕海英[1] 邱亚利[1] 

机构地区：[1]福建中医药大学药学院,福州350122

出　　处：《中国现代应用药学》2014年第1期44-48,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金项目(81202914);福建省科技计划项目(2011Y0035);福建省教育厅(JA12171)

摘　　要：目的 制备石杉碱甲单克隆抗体,分析其免疫学特性,建立蛇足石杉中石杉碱甲ELISA检测方法。方法 用石杉碱甲人工抗原免疫Balb/c小鼠, 采用杂交瘤技术筛选获取4株单克隆抗体3E02、4B01、5D03和4C04,对单克隆抗体纯度、亚型、灵敏度、特异性等进行鉴定,建立ELISA检测方法。结果 4株抗体亚型分别为IgG2b,IgG2b,IgG1和IgG1。ELISA检测方法的标准曲线方程为Y = 0.3736X - 0.236,r=0.9965,线性检测范围5-1000 ng/mL,检测限7.361 ng/mL。石杉碱甲的回收率在96.6-103.8%之间,RDS 0.68%（n=6）。结论 ELISA方法快速灵敏,可用于蛇足石杉中石杉碱甲的含量分析测定。Objective Preparation and characterization of monoclonal antibodies against Huperzine A, and establishing a determination method for HupA in Huperzia serrata.Methods：BALB/c mice were immunized by the conjugation of HupA–BSA successfully, and monoclonal antibodies were generated by hydridoma technique. Then hybridomas were cloned by the limiting dilution method and four hybridomas producing McAb reactive to HupA named 3E02, 4B01, 5D03 and 4C04 were obtained, a enzyme-linked immunosorbent assay for HupA was established successfully. Result： McAb isotype were IgG2b, IgG2b, IgG1 and IgG1.The ELISA standard curve was Y=0.363X-0.2377, r=0.9920, and the range of linearity was 5-1000 ng/mL. The recovery was 96.6-103.8%, RDS 0.68%（n=6）. Conclusion： The ELISA method provided a rapid,sensitive and accurate procedure for the determination of HupA in Huperzia serrata samples.

关 键 词：石杉碱甲 单克隆抗体 ELISA 蛇足石杉 

分 类 号：R962[医药卫生—药理学]