ENZYME IMMUNOASSAY BY CHEMILUMINESCENCE METHOD

摘要

PURPOSE:To make it possible to perform the micro-quantitative determination of the physiologically active materials in all vital fluids with high sensitivity by making enzyme immunity measurement by using the enzyme which generates hydrogen peroxide and measuring the activity of the enzyme by a prescribed chemiluminescence method. CONSTITUTION:The antibody Ab which conjugates specifically with the physiologically active material Ag in the vital fluid is conjugated with a solid phase and a sample contg. an unknown quantity of Ag is brought into reaction with the solid phase. the complex which is Ab-Ag-Ab-enzyme is formed on the solid phase when an enzyme- labeled antibody formed by labeling enzyme which generates hydrogen peroxide such as glucose oxidase with Ab is brought into reaction therewith. The quantity of the Ag is proportional to the quantity of the enzyme. The substrate of enzyme is added to such solid phase to generate H2O2 and an oxydation catalyst and a non-fluorescent material having oxidizability are added to the reactive liquid thereof to convert the non-fluorescent material to a fluorescent material. An inhibitor is then added to the reactive liquid to inhibit the oxidation catalyst and oxalic diester and H2O2 are added to the liquid so that the liquid emits light. The quantity of the emitted light is proportional to the quantity of the labeled enzyme, i.e., Ag and the quantity of the Ag is measured.