Production of specific monoclonal antibody against marker compound, hesperidin, geniposide, and saikosaponin b₂ for the quality control of crude drugs

Quality control of crude drugs based on marker compound is crucial as the quality directly affects efficacy of the treatment. So far, we have focused on plant secondary metabolites, and developed immunoassays for the quality control of crude drugs because highly sensitive, selective and efficacy quantitative/qualitative analysis can be carried out with immunoassay even though the sample is plant-derived sample which contains abundant secondary metabolites as well as primary metabolites, such as carbohydrates, lipids and amino acids.

In this study, we have produced three monoclonal antibodies (mAbs) for hesperidin (HP), geniposide (GP), and saikosaponin b₂ (SSb₂) to develop quality assessment method for Citrus unshiu peel, Gardenia fruit, and Bupleurum root, respectively, which were used in Kampo medicines. The marker compounds, HP, GP and SSb₂ were individually conjugated with bovine serum albumin to elicit an immunogenicity. Hybridomas secreting mAb were generated by fusing splenocytes and mice myeloma cells, SP2/0, and specific mAb to each compound was screened by indirect competitive enzyme-linked immunosorbent assay (icELISA). Characterisation of three purified mAbs by icELISA revealed that they were specific to each marker compound with high sensitivity, in which the limit of detection for HP, GP and SSb₂ exhibited were 1.12 ng/mL 1.13 μg/mL, and 2.13 ng/mL, respectively. These results indicated that the mAbs can be applied to quality control for the crude drugs containing HP, GP and SSb₂.

Production and characterisation of three mAbs, and their application to icELISA, will be presented in this conference.

 

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Conflict of Interest

The authors declare no conflict of interest.

Publication History

Article published online:
16 November 2023