Development of a solid-phase chemiluminescence immunoassay for plasma progesterone.
ABSTRACT A solid-phase immunoassay procedure for the determination of progesterone in human plasma is described, which utilizes chemiluminescence as the end-point. A progesterone-isoluminol conjugate serves as the chemiluminescent marker. An IgG fraction of antiserum to progesterone-11 alpha-hemisuccinate bovine serum albumin is passively adsorbed to the walls of Lumacuvettes P polystyrene test tubes. After the binding reaction, the cuvettes are washed. The light yield of the bound conjugate upon oxidation with a H2O2-microperoxidase system at pH 13 decreased with increasing free progesterone concentration in a log-linear manner over the range 15-1000 pg/tube. The chemiluminescence immunoassay is comparable to radioimmunoassay with regard to sensitivity, specificity, precision and accuracy. The assay offers the advantages of speed and ease of automation. In addition, this method does not involve the use of radioisotopes or of a centrifugation step.
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ABSTRACT: Although radioimmunoassay procedures have a number of advantages they do pose problems, especially with regard to the disposal of radioactive waste. Alternative methods, such as chemiluminescence, enzyme, fluorescence, or spin immunoassays, have been tested with the aim of replacing radioactive labels without loss of sensitivity, precision and accuracy.TrAC Trends in Analytical Chemistry 06/1983; 2(6):132-136. · 6.61 Impact Factor