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Single-type reporter multiplexing with A single droplet through bead-based digital microfluidics.

Meng-Shiue Lee1, Yen-Chia Chang2, Hong-Yuan Huang3

  • 1Department of Mechanical Engineering, National Yang Ming Chiao Tung University, Hsinchu 30010 Taiwan; Institute of Pharmacology, National Yang Ming Chiao Tung University, Taipei 11221, Taiwan.

Journal of Pharmaceutical and Biomedical Analysis
|June 19, 2022
PubMed
Summary

This study introduces single-type reporter multiplexing (STRM) for droplet microfluidics, enabling detection of multiple analytes from a single droplet using minimal sample volume and a single reporter type for enhanced biomedical diagnosis.

Keywords:
Biomedical diagnosticDigital microfluidicsMultiplex immunoassay

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Microfluidics

Background:

  • Droplet-based microfluidics offer advantages for biomedical diagnosis with limited sample volumes.
  • Multiplexing analytes typically requires larger sample volumes or advanced spectral detection methods.
  • Previous work achieved dual-analyte detection using different fluorescence wavelengths, limited by spectral resolution.

Purpose of the Study:

  • To develop a novel multiplexing approach for droplet microfluidics using a single sample droplet.
  • To enable detection of multiple analytes using a single-type reporter, overcoming spectral limitations.
  • To demonstrate the feasibility of single-type reporter multiplexing (STRM) for biomedical assays.

Main Methods:

  • Developed a bead-based digital microfluidic immunoassay.
  • Implemented a single-type reporter multiplexing (STRM) strategy by partitioning single droplets into sub-assays.
  • Validated the method for detecting human Interleukin-1 beta (IL-1β) and human Tumor Necrosis Factor-alpha (TNF-α).

Main Results:

  • Achieved multiplexed detection of two analytes (human IL-1β and human TNF-α) from a single 520 nL sample droplet.
  • Demonstrated standard curves for both analytes.
  • Reported low limits of detection (LOD) of 1.14 pg/mL for human IL-1β and 0.97 pg/mL for human TNF-α.
  • Completed the on-chip process in under 50 minutes.

Conclusions:

  • The proposed bead-based digital microfluidic immunoassay successfully enables multiplexed analyte detection from a single droplet using STRM.
  • This method overcomes spectral limitations and reduces sample volume requirements for multiplexed assays.
  • STRM offers a novel and efficient approach for sensitive, multi-analyte detection in biomedical diagnostics.