Multiplexed sandwich immunoassays using flow-injection electrochemiluminescence with designed substrate spatial-resolved technique for detection of tumor markers.

Convenient sensor array for simultaneous multi-analyte testing was increasingly needed in clinical diagnosis. A novel electrochemiluminescence (ECL) immunosensor array for the sequential detection of multiple tumor markers was developed by site-selectively immobilizing multiple antigens on different electrodes. Disposable indium tin oxide (ITO) glass array was employed as detection platform. With a sandwich-type immunoassay format, the amount of carbon dots coated silica (SiO(2)@C-dots) labeled antibodies increased with the increment of antigens in the samples. The ECL signal from different immunosensors was collected in turn by a photomultiplier (PMT) with the aid of a home-made potential transformer equiped with a home-made multiplexed-switch. Using carcino embryonic antigen (CEA), prostate specific antigen (PSA) and α-fetoprotein (α-AFP) as model analytes, the proposed immunoassay exhibited excellent precision and sensitivity. For all three analytes, the relative standard deviations (RSDs) for six times detection were lower than 7.1% and the detection limits were in the range of 0.003-0.006 ng mL(-1). The results for real sample analysis demonstrated that the newly constructed immunosensor array provided a rapid, simple, simultaneous multi-analyte immunoassay with high throughput, cost-effective and sufficiently low detection limits for clinical applications. Importantly, the novel individually addressable immunosensor array for multi-analyte immunoassay by introducing the ECL readout mechanism with the aid of the home-made potential transformer and multiplexed-switch could be a useful supplement to commercial assay methods in clinical chemistry.