Abstract: Additives capable of controlling the reaction rate between substrate and enzyme to form a dye in enzyme assays with absorbance, chemiluminescence, and fluorescence are effective tools for optimizing enzyme assays. A dye can be formed rapidly from the reaction between enzyme and substrate in the presence of a catalyst. Using relatively high concentration of dye formed from the rapid reaction, trace levels of analytical materials can be quantified using absorbance, chemiluminecence and fluorescence detection. A dye can be formed from a relatively slow reaction between enzyme and substrate in the presence of a surfactant such as Triton X-100 and ?-cyclodextrin. Using relatively low concentration of dye from the slow reaction, a high concentration of analytical material can be quantified without any dilution using absorbance, chemiluminescence, and fluorescence.

Abstract: A chemiluminescent enzyme immunoassay method for quantifying antigen or antibody using 1,1?-oxalyldiimidazole (ODI) derivative or 1,1?-oxalyldisodium benzoate (ODB) derivative chemiluminescence (CL) detection was developed. Also, various enzymes were quantified using ODI derivative or DOB derivative CL detection. Fluorescent compound formed from a substrate (non-fluorescent compound) through the enzyme assay methods emitted CL when the fluorescent compound received energy from high-energy intermediate formed in ODI derivative or ODB derivative CL reaction.