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.
