Abstract
A microtitre plate enzymeimmunoassay (EIA) for the determination of estradiol-17β (E2β)in bovine plasma was established. A homologous system was applied to this EIA. Antiserum against E2β-17-hemisuccinate bovine serum albumin was raised in rabbit. E2β-17-hemisuccinate peroxidase was used as a steroid-enzyme conjugate. Plasma E2β was extracted by diethyl ether and separated columun chromatography before EIA. Rabbit antiserum was used at 1:100,000 dilution for EIA. The sensitivity of the EIA was <0.25 pg/well. Out of the 12 steroids tests, high cross-reactivity was found in estrone (100%)and no cross-reactivity was found in C19 and C21 steroids. Recovery rates of E2β added to bovine plasma were 93.94-118.06% (mean=105.13%). Inter-and intra-assay coefficient of variations (CV) were 6.70-10.03% and 5.54-14.40%, respectivily. These results indicate that the EIA can be applied with sufficient sensitivity and precision for the routine analysis of bovine plasma E2β concentration.
