Analytical Sciences

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

Kazuhiko TSUKAGOSHI, Kazumasa TSUGE, and Riichiro NAKAJIMA

Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University, Kyotanabe 610-0321, Japan

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.

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