Development of an Assay System for Saikosaponin a Using Antisaikosaponin a Monoclonal Antibodies(Pharmacognosy)

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概要

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• For immunization, saikosaponin a (SSa) was conjugated with bovine serum albumin (BSA). The hapten number in an antigen conjugate was determined to be eleven by matrix-assisted laser adsorption/ionization time-of-flight mass spectrometry (MALDI-TOF Mass). Hybridomas secreting monoclonal antibodies (MAb) against SSa were produced by fusing splenocytes immunized with SSa-BSA conjugate and a hypoxanthine-aminopterin-thymidine-sensitive (HAT) mouse myeloma cell line, P3-X63-Ag8-653. A high specific MAb against SSa was selected from hybridomas using enzyme-linked immunosorbent assay (ELISA) analysis. Weak cross-reactivities occurred with saikosaponin c, b2 and d, which are stereochemical and/or functional isomers of SSa, but no crossreactivities were observed with other related steroidal glycosides. The full range of the assay extends 26 ng/ml to 1.5μg/ml of SSa. Good correlation of SSa concentrations in a crude extract of Bupleuri radix between ELISA and HPLC methods was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution. The newly established ELISA has been applied for the quantitative assay of SSa in the Bupleuri radix and the Kampo medicines (TCM) prescribed with Bupleuri radix.

• 公益社団法人日本薬学会の論文
• 2004-01-01

著者

• Tanaka Hiroyuki

  Department Of Cardiology Kurashiki Central Hospital

• Zhu Shu-hang

  Department Of Pharmacognosy Graduate School Of Pharmaceutical Sciences Kyushu University

• Tanaka Hiroyuki

  Graduate School of Pharmaceutical Sciences, Kyushu University

• Zhu Shu-hang

  九州大学 大学院薬学府生薬学

• Shoyama Yukihiro

  Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University

• SHIMOKAWA Shinichi

  Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University

• SHIMOKAWA Shin-ichi

  Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University

• Shimokawa Shin-ichi

  Department Of Pharmacognosy Graduate School Of Pharmaceutical Sciences Kyushu University

• Tanaka H

  Department Of Medicinal Plants Breeding And Pharmacognosy Graduate School Of Pharmaceutical Sciences

• Tanaka Hiroyuki

  Department Of Medicinal Plants Breeding And Pharmacognosy Graduate School Of Pharmaceutical Sciences

• Shoyama Y

  Department Of Pharmacognosy Faculty Of Pharmaceutical Sciences Nagasaki International University

• Shoyama Yukihiro

  Department Of Pharmacognosy Graduate School Of Pharmaceutical Sciences Kyushu University

• Tsukikawa Hiroshi

  Department Of Pharmacognosy Graduate School Of Medical Science

• Shoyama Yukihiro

  Department Of Forestry Faculty Of Agriculture Kyushu University

• Tanaka H

  Department Of Medicinal Plants Breeding And Pharmacognosy Graduate School Of Pharmaceutical Sciences

• Tanaka Hiroyuki

  Department Of Anesthesia And Clinical Research Institute National Hospital Organization Kyushu Medic

• SHOYAMA Yukihiro

  Department of Chemo-Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Kyushu University

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