抄録
We developed a time-resolved fluoro-immunoassay (TR-FIA) for choriogenin (Cg), a precursor of vitelline envelope protein, by using a specific antibody against Cg in marbled sole Pleuronectes yokohamae. Two major proteins, high- and low molecular-weight vitelline envelope proteins (VeH, 61 kDa and VeL, 37 kDa), constituted the vitelline envelope of the mabled sole. We purified Cg (61 kDa) - which showed common antigenicity with VeH ? from the serum of estrogen-treated marbled sole by a combination of ion-exchange and hydroxylapatite chromatography. The specific antibody against Cg immunostained 170 and 61 kDa bands of vitelline envelope and 61 kDa band of the serum of estrogen-treated marbled sole. The dilution curve of purified Cg was parallel to that of the serum of estrogen-treated marbled sole. The standard curve of TR-FIA was almost linear from 61 pg/mL to 31.3 ng/mL. The intra- and inter-assay variations were less than 5.1 and 12.8 %, respectively. The high sensitivity of TR-FIA will be useful in the detection of effect by environmental estrogens.
