一种基于酶联免疫吸附测定（ELISA）的新型药理活性柴胡皂苷分析方法。

A novel analytical ELISA-based methodology for pharmacologically active saikosaponins.

作者信息

Zhu S, Shimokawa S, Shoyama Y, Tanaka H

机构信息

Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan.

出版信息

Fitoterapia. 2006 Feb;77(2):100-8. doi: 10.1016/j.fitote.2005.11.005. Epub 2006 Jan 10.

DOI:10.1016/j.fitote.2005.11.005

PMID:16376495

Abstract

A competitive enzyme-linked immunosorbent assay (ELISA) for saikosaponins was established using monoclonal antibody (MAb) 3G10. Hybridoma 3G10 prepared by fusing splenocytes immunized with saikosaponin a-BSA (SSa-BSA) conjugate and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3-X63-Ag8-U1, secreted monoclonal antibodies with wide cross-reactivity to saikosaponins including saikosaponin b(2) (SSb(2)), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 mug /ml to 2.3 mug/ml of SSa, successfully detected total saikosaponins in Bupleuri radix and Kampo medicines prescribed with Bupleuri radix. Good correlation between ELISA and HPLC analyses of total saikosaponin in a crude extract of Bupleuri radix was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution.

摘要

使用单克隆抗体（MAb）3G10建立了一种用于柴胡皂苷的竞争性酶联免疫吸附测定（ELISA）方法。通过将用柴胡皂苷a-牛血清白蛋白（SSa-BSA）偶联物免疫的脾细胞与次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷（HAT）敏感的小鼠骨髓瘤细胞系P3-X63-Ag8-U1融合制备的杂交瘤3G10，分泌对包括柴胡皂苷b(2)（SSb(2)）、c（SSc）和d（SSd）在内的柴胡皂苷具有广泛交叉反应性的单克隆抗体，这些都是SSa的立体和/或区域异构体。该方法在SSa的有效测量范围为0.6μg/ml至2.3μg/ml时，成功检测了柴胡及含柴胡的汉方制剂中的总柴胡皂苷。在用温和碱性溶液处理使酰基柴胡皂苷水解后，ELISA与柴胡粗提取物中总柴胡皂苷的HPLC分析之间获得了良好的相关性。