甘草酸多克隆抗体的制备及ELISA方法建立  

Preparation of Glycyrrhizinic Acid Polyclonal Antibody and Development of ELISA for Glycyrrhizinic Acid Detection

作　　者：李刚[1,2] 赵静[2] 何素平[2] 邓艾兴[2] 李召虎[2] 何钟佩[2] 赵环环[2] 王保民[2] 

机构地区：[1]吉林省农业科学院农业环境与资源研究中心,吉林长春130033 [2]中国农业大学农学与生物技术学院,北京100094

出　　处：《时珍国医国药》2011年第12期2956-2958,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家"863"高技术研究发展计划(子课题)(No.2008AA10Z426)

摘　　要：目的制备甘草酸多克隆抗体及建立甘草酸酶联免疫吸附测定(ELISA)方法。方法通过碳化二亚胺法活化甘草酸与载体蛋白BSA联结制备免疫抗原,并免疫Balb/c小鼠,获得了甘草酸多克隆抗体并建立了甘草酸ELISA检测方法。用所建立的甘草酸ELISA检测方法对4个甘草样品进行了检测并用HPLC法加以验证。结果抗体效价>3.2×106,与甘草次酸的交叉反应率为24.2%。ELISA检测范围为3～120 ng/ml,IC50为20 ng/ml。添加实验的回收率为103.5%～108%。结论 ELISA方法测定的数据与HPLC方法测定的数据有很好的相关性(R2=0.990 1)。Objective To produce polyclonal antibodies（PcAbs） and to develop an enzyme-linked immunosorbent assay（ELISA） for glycyrrhizinic acid. Methods Glycyrrhizinic acid（GA） was covalently conjugated to carrier protein（bovine serum albumin,BSA） to prepare immunogen by the carbodiimide（CDI） method.GA-BSA conjugate was used to immunize Balb/c mice to produce polyclonal antibodies（PcAbs）.An enzyme-linked immunosorbent assay（ELISA） for glycyrrhizinic acid was developed. Results The antibody titer was found as higher than 3.2×106.Cross-reactivity with glycyrrhetic acid was 24.2%.The detection range was 3～120 ng/ml and the value of IC50 was 20 ng/ml.Recoveries of GA spikes were between 103.5% and 108%. Conclusion Glycyrrhizinic acid content of four powdery licorice root samples was determined by ELISA and validated by HPLC.Results show that data determined by the ELISA are highly correlated with those by HPLC（R2=0.990 1）.

关 键 词：甘草酸 抗体 酶联免疫吸附测定法 

分 类 号：R283[医药卫生—中药学]