芦丁完全抗原的合成鉴定及免疫原性分析(4)
(1.Department of Pharmaceutics， Hunan University of Traditional Chinese Medicine， Changsha 410208， China；
2. Property and Pharmacodaynamic Key Laboratory of Traditional Chinese Medicine， State Administration of Traditional
Chinese Medicine， Changsha 410208， China)
[Abstract] Objective： Synthesis and identification of complete antigen of rutin， the traditional Chinese medicine active ingredient， and develop rapid detection of rutin using enzyme-linked immunoassay method (ELISA). Immunogenicity of the complete antigen 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 bovine serum protein. The titer of rutin antibody is 1∶4 000. The sensitivity of IC50 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.
[Key words] rutin； complete antigen； synthesis； identification； immunogenicity
doi：10.4268/cjcmm20130321
[责任编辑 张宁宁], 百拇医药(包小燕，贺福元，曾姣丽，夏赞韶，邓俊林，孙青辉，雷虹，杜玉然)
2. Property and Pharmacodaynamic Key Laboratory of Traditional Chinese Medicine， State Administration of Traditional
Chinese Medicine， Changsha 410208， China)
[Abstract] Objective： Synthesis and identification of complete antigen of rutin， the traditional Chinese medicine active ingredient， and develop rapid detection of rutin using enzyme-linked immunoassay method (ELISA). Immunogenicity of the complete antigen 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 bovine serum protein. The titer of rutin antibody is 1∶4 000. The sensitivity of IC50 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.
[Key words] rutin； complete antigen； synthesis； identification； immunogenicity
doi：10.4268/cjcmm20130321
[责任编辑 张宁宁], 百拇医药(包小燕，贺福元，曾姣丽，夏赞韶，邓俊林，孙青辉，雷虹，杜玉然)