Development of an immune microanalysis system by use of peroxyoxalate chemiluminescence detection.
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.
Tsukagoshi
Kazuhiko
K
Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University, Japan.
Tsuge
Kazumasa
K
Nakajima
Riichiro
R
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001667:Binding, Competitive; D046209:Electrophoresis, Microchip / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry; D005898:Glass / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D008163:Luminescent Measurements / Q000379:methods*; D046208:Microchip Analytical Procedures / Q000379:methods*; D009363:Neoplasm Proteins / Q000097:blood / Q000737:chemistry / Q000276:immunology; D010070:Oxalates / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D012709:Serum Albumin / Q000032:analysis / Q000737:chemistry / Q000276:immunology
D000906:Antibodies; C518288:FITC-albumin; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D010070:Oxalates; D011506:Proteins; D012709:Serum Albumin; C030193:immunosuppressive acidic protein; C054009:bis(4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl)oxalate; C044622:peroxyoxalate; D006861:Hydrogen Peroxide; D016650:Fluorescein-5-isothiocyanate