Measurement of steroids by enzymeimmunoassay: An environmentally
friendly alternative to radioimmunoassay
K.M. Henderson, A.H.M. Hardie, M. Camberis
AgResearch, Wallaceville Animal Research Centre, P O Box 40063,
Upper Hutt, New Zealand.
Proceedings of the New Zealand Society of Animal Production 1994,
54: 55-58
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.
Keywords: NZSAPAB;
enzymeimmunoassay; radioimmunoassay; oestradiol-17b; progesterone;
granulosa cells.
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Last Updated 25-01-1997
