Characterization and validation of fragment antigen-binding (Fab)
antibody-based immunoassay for deoxymiroestrol, a potent phytoestrogen
from Pueraria candollei
Deoxymiroestrol is the most potent phytoestrogen in chromenes group that
has been found in Pueraria candollei, Thai name known as Kwao Krua Khao.
Several studies reported estrogenic activity of P. candollei in order to
using as hormone replacement therapy for postmenopausal women.
Previously, specific determination of deoxymiroestrol content by
enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody
(pAb) have been reported. However, production of pAb has limitation and
variability from different batches. Therefore, in this study, we
established quantitative method for determination of deoxymiroestrol
using fragment antigen-binding (Fab) antibody-based immunoassay. The
developed immunoassay has specificity to deoxymiroestrol with a
calibration range of 15.6-1000 ng mL-1. Precision including intra-assay
and inter-assay are 1.48-7.11 and 0.58-9.31%, respectively. Accuracy of
the assay showed in recovery between 99.77-101.61% when spike
deoxymiroestrol standard into the samples. The limit of detection (LOD)
is 30.80 ng mL-1. Comparation antibody-based immunoassay for
determination of deoxymiroestrol using Fab with pAb was represented
consistency (R2 = 0.9807) when analysis roots bark of Pueraria candollei
from difference areas. Therefore, this development assay can apply to
determine deoxymiroestrol content in the plant samples.