RRC ID 77006
Author Ochi A, Fujii S, Ohta T, Uto T, Nuntawong P, Tanaka H, Morimoto S, Sakamoto S.
Title Highly sensitive indirect competitive enzyme-linked immunosorbent assay based on a monoclonal antibody against saikosaponin b2 for quality control of Kampo medicines containing Bupleuri radix.
Journal J Nat Med
Abstract Saikosaponins are naturally occurring oleanane-type triterpenoids that are found in Bupleuri radix (root of Bupleurum falcatum) and exhibit a broad biological activity spectrum. Saikosaponin b2 (SSb2) is the main saikosaponin in Kampo medicine extracts and is a designated quality control marker for the same in the Japanese Pharmacopeia. Although some monoclonal antibodies (mAbs) against saikosaponins have been produced to evaluate the quality of Bupleuri radix and related products, anti-SSb2 mAbs have not been used to quantify SSb2 in Kampo medicines. To address this knowledge gap, we herein established a new hybridoma cell line secreting a highly specific anti-SSb2 mAb and developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on this mAb for the detection of SSb2 in Bupleuri radix-containing Kampo medicines. The generated SSb2-recognized mAb exhibited high specificity to SSb2 in icELISA. The developed assay featured high sensitivity (linearity range = 1.95-125 ng/ml), accuracy, precision and reproducibility (coefficient of variation < 5%), and the thus determined SSb2 contents were strongly correlated with those obtained using liquid chromatograph-mass spectrometer. These results suggest that the anti-SSb2 mAb-based icELISA method can be used for the quality control and standardization of Kampo medicines containing Bupleuri radix.
Volume 78(1)
Pages 160-168
Published 2024-1-1
DOI 10.1007/s11418-023-01753-3
PII 10.1007/s11418-023-01753-3
PMID 37804411
MeSH Antibodies, Monoclonal Enzyme-Linked Immunosorbent Assay Medicine, Kampo Oleanolic Acid* / analogs & derivatives Quality Control Reproducibility of Results Saponins* / analysis
IF 2.055
Resource
Human and Animal Cells SP2/0-Ag14(RCB0209)