TY - JOUR
T1 - A novel analytical ELISA-based methodology for pharmacologically active saikosaponins
AU - Zhu, S.
AU - Shimokawa, S.
AU - Shoyama, Y.
AU - Tanaka, H.
N1 - Funding Information:
This paper was supported in part by a Grant-in-Aid (No. 08457586 for Y.S.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the research fund of Kyushu University Foundation, and the research fund of the Japan Kampo Medicine Manufacturers Association from Japan Kampo Medicine Manufacturers.
PY - 2006/2
Y1 - 2006/2
N2 - 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 b2 (SSb2), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 μg /ml to 2.3 μg/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.
AB - 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 b2 (SSb2), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 μg /ml to 2.3 μg/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.
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U2 - 10.1016/j.fitote.2005.11.005
DO - 10.1016/j.fitote.2005.11.005
M3 - Article
C2 - 16376495
AN - SCOPUS:32344432081
SN - 0367-326X
VL - 77
SP - 100
EP - 108
JO - Fitoterapia
JF - Fitoterapia
IS - 2
ER -