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High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity.

Connie Wu1,2, Tyler J Dougan1,2,3, David R Walt1,2

  • 1Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, United States.

ACS Nano
|January 14, 2022
PubMed
Summary

We developed MOSAIC, a digital ELISA platform, achieving attomolar sensitivity for detecting rare protein biomarkers. This technology offers enhanced throughput and multiplexing for improved clinical diagnostics and biomarker discovery.

Keywords:
biomarkersdiagnosticsdigitalimmunoassaysingle moleculeultrasensitive protein detection

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Biomarker Discovery

Background:

  • Measuring low-abundance proteins in biological fluids is crucial for clinical diagnostics but challenging with conventional methods.
  • Digital ELISA technologies offer increased sensitivity but often lack the required performance for rare biomarkers and require specialized setups.
  • Existing digital ELISA platforms face limitations in sensitivity, throughput, and ease of implementation for widespread clinical use.

Purpose of the Study:

  • To develop a highly sensitive and streamlined digital ELISA platform for detecting low-abundance proteins.
  • To overcome the limitations of current digital ELISA technologies in terms of sensitivity, workflow, and instrumentation.
  • To enable ultrasensitive detection and multiplexed measurements of protein biomarkers for clinical applications.

Main Methods:

  • Development of the Molecular On-bead Signal Amplification for Individual Counting (MOSAIC) platform.
  • Utilizing a rapid, automatable flow cytometric readout for high-throughput analysis.
  • Employing tunable assay bead numbers for enhanced signal-to-background ratios and precision.

Main Results:

  • MOSAIC achieves low attomolar limits of detection, an order of magnitude more sensitive than existing methods.
  • The platform demonstrates high sampling efficiencies for rare target molecules.
  • Successful application in detecting low-abundance cytokines in saliva and multiplexed protein analysis in plasma and saliva.

Conclusions:

  • MOSAIC provides ultrasensitive protein detection with attomolar sensitivity and high throughput.
  • The platform's versatility supports higher-order multiplexing with femtomolar sensitivity, expanding diagnostic capabilities.
  • MOSAIC offers accessible and versatile ultrasensitive detection, potentially accelerating biomarker discovery and clinical diagnostics.