Monoclonal antibody production against majonoside-R2, the major bioactive compound of Vietnamese ginseng

Panax vietnamensis, or Vietnamese ginseng (VG), is a Panax sp. that was a plant discovered and proven to possess multifunctional biological activities. Chemical constituents of VG have been studied, and majonoside-R2 (MR2) has been found as a major ocotillol-type saponin containing around 5.3% in addition to protopanaxadiol-type and protopanaxatriol-type saponins. As with other Panax spp., VG is used for numerous treatment indications, including immune system enhancement, inflammatory reduction, anti- cancer, memory loss protection, and aging prevention; furthermore, it has been reported as a dominant anti-myocardial ischemia. Therefore, MR2 became a consequential bioactive compound in VG that needs to be investigated. In previous studies, HPLC and HPLC/evaporative light scattering detector were the only methods performed in order to detect the ocotillol saponin. The difficulty in detecting this saponin-type was caused by the lack of appropriate chromophores for UV absorbance. Thus, immune-specific detection using monoclonal antibodies (mAb) was developed for the purpose of providing the analytical benefit of MR2 in VG. In this study, BALB/c mice were immunised with MR2-BSA conjugates prepared by oxidative amination using sodium periodate. After cell fusion of splenocytes with SP2/0 myeloma cells, three outstanding hybridoma candidates secreting anti-MR2 antibodies were screened on the basis of specificity. Finally, the best hybridoma cells secreting highly specific MR2 mAb was chosen for further expansion. In conclusion, MR2 mAb was successfully obtained as a useful tool for nanogram-level quantitative analysis detection for MR2 (3.90 – 125 ng/mL) using enzyme-linked immunosorbent assay.

Publication History

Article published online:
16 November 2023