芦丁完全抗原的合成鉴定及免疫原性分析  

Synthesis and identification of rutin complete antigen and analysis its immunogenicity

作　　者：包小燕[1] 贺福元[1,2] 曾姣丽[1] 夏赞韶[1] 邓俊林[1] 孙青辉[1] 雷虹[1] 杜玉然[1] 

机构地区：[1]湖南中医药大学药学院,湖南长沙410208 [2]中药药性与药效国家中医药管理局重点实验室,湖南长沙410208

出　　处：《中国中药杂志》2013年第3期397-401,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81073142);湖南省教育厅重点项目(09A068);湖南省自然基金重点项目(11JJ2055)

摘　　要：目的:合成中药活性成分芦丁的完全抗原并进行鉴定及免疫原性研究,同时建立快速检测芦丁的酶联免疫吸附(ELISA)分析方法。方法:采用高碘酸钠法制备芦丁完全抗原,用紫外扫描和SDS-PAGE的方法对其进行鉴定,以此抗原免疫健康雄性新西兰大白兔,制备抗血清。结果:紫外分析表明合成的完全抗原偶联比为13∶1,SDS-PAGE显示完全抗原与牛血清蛋白或卵清蛋白相比后移了。免疫新西兰大白兔得到特异针对芦丁的抗血清,芦丁抗体的效价为1∶4 000。经方法学考察,灵敏度IC50为5.37 mg.L-1,最低检测限为1 mg.L-1,平均添加回收率为102%,批内和批间RSD均小于10%,抗体与其他类似物的交叉反应率小于1%。结论:成功的合成了具有较好的免疫原性的芦丁完全抗原,并成功的建立了快速检测芦丁的酶联免疫吸附(ELISA)方法。Objective： Synthesis and identification of complete antigen of rutin, the traditional Chinese medicine active ingre- dient, and develop rapid detection of rutin using enzyme-linked immunoassay method （ELISA）. Immunogenicity of the complete anti- gen was also studied. Method： Prepare the complete antigen by sodium periodate solution and identified by UV scanning and SDS- PAGE test. Male New Zealand white rabbits were immunized by the antigen to obtain the antiserum. Result ： The results of UV analysis showed that the coupling ratio of complete antigen is 13： 1. SDS-PAGE display of the artificial antigen was delayed compared with bo- vine serum protein. The titer of rutin antibody is 1 ： 4 000. The sensitivity of ICs0 was 5.37 mg L- 1, the lowest detection limit was 1 mg L-1 ,the average recovery was 102%, the intra and interspecific RSD were less than 10% , cross-reactivity rate of antibodies and other analogs were less than 1%. Conclusion： Rutin complete antigen was synthesized successfully, and the rapid detection of rutin by ELISA method was successfully established.

关 键 词：芦丁 完全抗原 合成 鉴定 免疫原性 

分 类 号：R284[医药卫生—中药学]