摘要
目的研究建立双黄连注射液(金银花、黄芩、连翘)中半抗原成分的间接ELISA检测方法。方法将黄芩苷(BAL)与牛血清白蛋白(BSA)偶联,制备完全抗原BAL-BSA,免疫家兔获得针对BAL的特异性抗体。以BAL为包被抗原,优化用于检测抗血清效价的间接ELISA法的各项实验条件,并采用优化后的ELISA法检测双黄连注射液中的半抗原成分。结果完全抗原BAL-BSA免疫获得的高效价兔抗血清可以很好地识别BAL;所建立的ELISA法检测结果与双黄连注射液中BAL的量、临床ADR之间存在一定的相关性。结论合成的BAL-BSA完全抗原具有良好的反应原性与免疫原性;双黄连注射液致敏的主要抗原决定簇与BAL的结构有关;所建立的间接ELISA可用于检测注射液中与BAL结构相关的微量抗原性成分。
AIM To establish an indirect enzyme-linked immunosorbent assay ELISA） for detecting hapten in Shuanghuanglian Injection （Lonicerae japonicae Flos, Sctttellariae Radix, Forsythiae Fructus ）. METHODS The complete antigen （BAL-BSA） was synthesized by coupled baicalin （BAL） to bovine serum albumin （BSA） u- sing activated ester method. Rabbits were immunized with BAL-BSA to produce antiserum against BAL. The opti- mal conditions of indirect ELISA for the determination of antisera titers were investigated. The hapten in Shuang- huanglian Injection was detected by the indirect ELISA. RESULTS The antisera immunized by BAL-BSA with high titer could well recognize BAL. And the results of the optimized indirect ELISA had a correlationship between the ADR in clinic and the content of BAL in Shuanghuanglian Injection. CONCLUSIONS The synthesized com- plete antigen （BAL-BSA） has good immunogenicity and reactogenicity. The major antigenic determination of Shuanghuanglian Injection is related to structure with BAL, and the established ELISA is able to detect and control the potential trace antigenic compositions.
出处
《中成药》
CAS
CSCD
北大核心
2013年第12期2678-2683,共6页
Chinese Traditional Patent Medicine
关键词
黄芩苷
双黄连注射液
半抗原
过敏反应
酶联免疫吸附分析
抗原决定簇
baicalin
Shuanghuanglian Injection
hapten
allergic reaction
enzyme-linked immunosorbent assay （ELISA）
antigenic determinant
