A novel analytical ELISA-based methodology for pharmacologically active saikosaponins.
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.
Zhu
S
S
Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan.
Shimokawa
S
S
Shoyama
Y
Y
Tanaka
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000961:Antilymphocyte Serum / Q000032:analysis / Q000096:biosynthesis; D029972:Bupleurum / Q000737:chemistry; D002138:Calibration; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003429:Cross Reactions; D004365:Drugs, Chinese Herbal / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006241:Haptens / Q000032:analysis; D006825:Hybridomas; D016014:Linear Models; D051379:Mice; D008807:Mice, Inbred BALB C; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification; D010936:Plant Extracts / Q000032:analysis* / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry; D012503:Saponins / Q000032:analysis* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013223:Statistics as Topic / Q000379:methods
D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; C051274:Bupleurum root extract; D004365:Drugs, Chinese Herbal; D006241:Haptens; D010936:Plant Extracts; D012503:Saponins; C461320:shosaiko-to; D009828:Oleanolic Acid; C025759:saikosaponin D