Development of an enzyme immunoassay for the determination of testosterone
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In this study a double antibody enzyme immunoassay for the direct determination of testosterone in plasma was developed. Testosterone 3-O-CMO was conjugated with horseradish peroxidase by the mixed anhydride method and the conjugate purified by column chromatography (sephadex G-25). Testosterone antibody was obtained by immunization of rabbits against Testosterone-3-O-CMO-BSA. Cross reactions of the antiserum against some steroids were found to be <0.01%. The detection limit of the assay was 2.5 pg/well and the working range of the standard curve was 0-20 ng/ml (0-200 pg/well). The recovery was found to be 98.8% after the addition of known amounts of testosterone to plasma samples. The inter-assay coefficient of variation was 12.6%. The described EIA offers a very economical alternative for the routine estimation of testosterone levels. Only minimal laboratory equipment is required and therefore the assay should be especially useful for research in animal science.
SourceTurkish Journal of Veterinary & Animal Sciences