Development of an Immune Microanalysis System by Use of Peroxyoxalate Chemiluminescence Detection

Abstract

We developed an immune microanalysis system incorporating chemiluminescence detection, where the peroxyoxalate chemiluminescence (CL) detection using bis[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate (TDPO)-hydrogen peroxide (H₂O₂)-fluorescein isothiocianate (FITC) reaction was newly adopted. The analysis system performed the following three processes on a microchip: immune reaction for high selectivity, electrophoresis for formation and transportation of the sample plug, and CL detection. The immune reaction was carried out using an antibody-immobilized glass bead. The glass bead was placed in one of the reservoirs in the microchip along with antigen (analyte) and a known amount of FITC-labeled antigen to set up a competitive immune reaction. The reactant after the immune reaction was fed electrophoretically into the intersection, resulting in a sample plug. The sample plug was then moved into another reservoir containing TDPO-H₂O₂ acetonitrile solution. At this point, CL detection was performed. The system described here was capable of determining human serum albumin or immunosuppressive acidic protein as a cancer marker in human serum.