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

Overview

Journal J Nat Med

Publisher Springer

Date 2023 Oct 7

PMID 37804411

Authors

Akihiro Ochi

Shunsuke Fujii

Tomoe Ohta

Takuhiro Uto

Poomraphie Nuntawong

Hiroyuki Tanaka

Satoshi Morimoto

Seiichi Sakamoto

Affiliations

Soon will be listed here.

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.

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