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Dielectrophoresis assisted high-throughput detection system for multiplexed immunoassays
Shih-Mo Yang1, Qiang Lin1, Hongbo Zhang2
1Biomedical Science and Technology Research Center, School of Mechatronic Engineering and Automation, Shanghai University, Shanghai, China.
Biosensors & Bioelectronics
|March 13, 2021
Summary
This study enhances digital enzyme-linked immunosorbent assay (ELISA) sensitivity by using electrokinetic effects to isolate single molecules for multiplexed cytokine detection. The method achieves pg/ml level detection for IL-2, IL-6, IL-10, and TNF-α.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Biochemistry
Background:
- Digital ELISA offers advantages for single-molecule detection, but sensitivity improvement is crucial.
- Current research focuses on enhancing the sensitivity of digital ELISA platforms.
- Simultaneous detection of multiple analytes remains a challenge in sensitive immunoassays.
Purpose of the Study:
- To develop a novel immunoassay method to improve the sensitivity and efficiency of simultaneous cytokine detection using digital ELISA.
- To apply electrokinetic effects for precise manipulation and isolation of encoded beads in microfluidic devices.
- To achieve high-sensitivity, multiplexed quantification of cytokines at the picogram per milliliter level.
Main Methods:
- Implemented a microfluidic device with a non-uniform electric field to induce dielectrophoresis (DEP) for bead manipulation and confinement.
- Utilized two wavelengths of excitation light for simultaneous detection of reporters on encoded beads.
- Employed total internal reflection to confine excitation light to the bottom slide for enhanced signal detection.
- Quantified cytokine concentrations by analyzing fluorescent intensity from individual beads after capture.
Main Results:
- Dielectrophoresis (DEP) increased the fill percentage of encoded beads in micro-wells from 10-20% to 60-80%.
- Achieved picogram per milliliter (pg/ml) level detection for four cytokines: IL-2, IL-6, IL-10, and TNF-α.
- Spike and recovery experiments demonstrated high capture efficiency, exceeding 70% for target molecules.
- Method reliability was validated through comparison with flow cytometry.
Conclusions:
- The application of dielectrophoresis (DEP) significantly enhances the sensitivity of digital ELISA for multiplexed cytokine detection.
- This electrokinetic-enhanced immunoassay enables rapid and sensitive quantification of multiple cytokines simultaneously.
- The developed platform shows promise for advancing sensitive single-molecule detection in various biological and clinical applications.
