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Updated: May 17, 2026
Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
Published on: June 2, 2023
Multiplexed sandwich immunoassays using flow-injection electrochemiluminescence with designed substrate
Yan Zhang1, Weiyan Liu, Shenguang Ge
1Key Laboratory of Chemical Sensing & Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, PR China.
Insights
A novel electrochemiluminescence (ECL) immunosensor array enables simultaneous detection of multiple tumor markers. This cost-effective, high-throughput method offers sensitive and precise results for clinical diagnosis.
Area of Science:
- Analytical Chemistry
- Biomedical Engineering
- Biosensors
Background:
- Clinical diagnosis requires convenient sensor arrays for simultaneous multi-analyte testing.
- Existing methods may lack the necessary throughput and cost-effectiveness for widespread clinical application.
Purpose of the Study:
- To develop a novel electrochemiluminescence (ECL) immunosensor array for sequential, simultaneous detection of multiple tumor markers.
- To establish a cost-effective, high-throughput immunoassay platform for clinical diagnosis.
Main Methods:
- Site-selective immobilization of multiple antigens on a disposable indium tin oxide (ITO) glass array.
- Sandwich-type immunoassay format utilizing carbon dots coated silica (SiO(2)@C-dots) labeled antibodies.
- Sequential ECL signal collection using a photomultiplier (PMT) with a custom potential transformer and multiplexed-switch.
Main Results:
- The immunosensor array demonstrated excellent precision (RSDs < 7.1%) and sensitivity for detecting carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), and α-fetoprotein (α-AFP).
- Detection limits were achieved in the range of 0.003–0.006 ng mL⁻¹.
- Real sample analysis confirmed the array's suitability for rapid, simultaneous multi-analyte immunoassay with high throughput and low detection limits.
Conclusions:
- The developed ECL immunosensor array offers a rapid, simple, and cost-effective solution for simultaneous multi-analyte immunoassay in clinical settings.
- This individually addressable immunosensor array represents a valuable supplement to existing commercial assay methods in clinical chemistry.
Abstract:
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.
