Abstract
Highly sensitive enzyme immunoassays using chemiluminescence reaction have been developed. Horseradish peroxidase and glucose oxidase were used as the labeling enzyme and conjugated with cortisol, dehydroepiandrosterone and its sulfate, 17α-hydroxyprogesterone, thyroxine, α-fetoprotein and insulin, respectively. Free and bound fractions after immune reaction, were separated by insoliblized antibodies or secondary antibodies.
The enxyme activities were measured by chemiluminescence reaction using luminol and hydrogen peroxide as substrates for horseradish peroxidase, and the glucose oxidase activity was also measured by luminol and potasium ferricyanide system/or trichlorophenol oxalate-fluorescence dye system after incubation with glucose. The faint chemiluminescence was measured by a photon counter or flow injection analysis system. Comparison of assay results obtained by radioimmunoassay and enzymeimmunoassay showed excellent agreement of results in all cases.
The detection limits of each substances were about 10-15-10-17 mol/assay tube. These enzymeimmunoassay based on chemiluminescence reaction were applicable to the routine determination of clinically important samples.
