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.
出处
《中国中药杂志》
CAS
CSCD
北大核心
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)
关键词
芦丁
完全抗原
合成
鉴定
免疫原性
rutin
complete antigen
synthesis
identification
immunogenicity
