A monoclonal antibody-based competitive ELISA for the determination of ruscogenin in Chinese traditional medicines and biological samples  

作　　者：XU Yu LIU Ji-Hua WANG Jing ZHANG Jian YU Bo-Yang 

机构地区：[1]Department of Complex Prescription of Traditional Chinese Medicine,China Pharmaceutical University [2]Key Laboratory of Modern Chinese Medicines,China Pharmaceutical University

出　　处：《Chinese Journal of Natural Medicines》2014年第10期794-799,共6页中国天然药物（英文版）

基　　金：supported by the Key Project of Chinese Ministry of Education(No.108071);the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD);the National Natural Science Foundation of China(No.81173471)

摘　　要：A competitive enzyme-linked immunosorbent assay(ELISA) was developed to determine ruscogenin(RUS) by using the monoclonal antibody(McAb). The monoclonal antibody against RUS, secreted from the established hybridoma cell lines, was identified as being of the IgG1 isotype. The McAb exhibited high specificity to RUS, showing a very slight cross reactivity with diosgenin(15.7%), and no cross-reactivity to sarsasapogenin, diammonium glycyrrhizinate, oleanolic acid and notoginsenoside R1. The established ELISA, at an IC50 value of 157.55 ng.mL-1 and a detection limit(IC20) of 20.57 ng.mL-1, was compared with HPLC analyses, and a good correlation between ELISA and HPLC-ELSD analyses of RUS in the extract of Radix Ophiopogonis was obtained. The experimental data indicated that the ELISA method exhibits more advantages over HPLC-ELSD, such as low detection limit, high specificity, low background, and no requirement for sample pre-treatment, and is more suitable for the determination of natural components in Chinese traditional medicines and in biological samples for pharmacokinetic studies.A competitive enzyme-linked immunosorbent assay（ELISA） was developed to determine ruscogenin（RUS） by using the monoclonal antibody（McAb）. The monoclonal antibody against RUS, secreted from the established hybridoma cell lines, was identified as being of the IgG1 isotype. The McAb exhibited high specificity to RUS, showing a very slight cross reactivity with diosgenin（15.7%）, and no cross-reactivity to sarsasapogenin, diammonium glycyrrhizinate, oleanolic acid and notoginsenoside R1. The established ELISA, at an IC50 value of 157.55 ng.mL-1 and a detection limit（IC20） of 20.57 ng.mL-1, was compared with HPLC analyses, and a good correlation between ELISA and HPLC-ELSD analyses of RUS in the extract of Radix Ophiopogonis was obtained. The experimental data indicated that the ELISA method exhibits more advantages over HPLC-ELSD, such as low detection limit, high specificity, low background, and no requirement for sample pre-treatment, and is more suitable for the determination of natural components in Chinese traditional medicines and in biological samples for pharmacokinetic studies.

关 键 词：EL1SA Monoclonal antibody Ruscogenim Radix Ophiopogonis PHARMACOKINETIC 

分 类 号：R917[医药卫生—药物分析学]