Enzymeimmunoassays (EIAs) for oestradiol-17b and progesterone were developed using antibody-coated microtitre plates, and steroid-horseradish peroxidase conjugates as steroid 'tracers'. The characteristics of standard curves and the precision of measurements were comparable between the EIAs and corresponding radioimmunoassays (RIAs). The applicability of the EIAs was demonstrated by using them to measure oestradiol-17b and progesterone concentrations in media sampled daily from cultures of bovine granulosa cells. The EIAs detected the characteristic switch in cellular steroidogenic capacity from oestradiol-17b to progesterone production typical of cultured granulosa cells. There was a significant linear correlation (P<0.001) between the EIAs and RIAs in the concentrations of steroids measured in granulosa cell culture medium by the two immunoassay formats. These findings may encourage other laboratories to consider converting "in-house" RIAs to EIAs, thereby eliminating the potential health and environmental hazards associated with the radioisotopes required for RIA.
Proceedings of the New Zealand Society of Animal Production, Volume 54, , 55-58, 1994
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