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Digital immunoassay for biomarker concentration quantification using solid-state nanopores.

Liqun He1, Daniel R Tessier1, Kyle Briggs1

  • 1Department of Physics, University of Ottawa, Ottawa, Canada.

Nature Communications
|September 10, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a digital immunoassay using DNA nanostructures and nanopores for precise protein biomarker quantification. This method enables accurate digital diagnostics for precision medicine, even in complex biological samples.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Molecular Diagnostics

Background:

  • Single-molecule counting offers unparalleled accuracy for biomarker concentration determination, driving the development of digital diagnostic platforms for precision medicine.
  • Solid-state nanopores, as fully electronic sensors with single-molecule sensitivity, are theoretically ideal for this application but face challenges in protein sensing.
  • Existing nanopore sensing methods struggle with specificity, sensitivity, and consistency when analyzing proteins in complex biological fluids.

Purpose of the Study:

  • To develop a robust digital immunoassay scheme for reliable protein concentration quantification in complex biofluids.
  • To overcome the limitations of solid-state nanopores in specificity, sensitivity, and consistency for protein detection.
  • To establish a proof-of-concept for a digital diagnostic approach using DNA nanostructures as protein proxies.

Main Methods:

  • A magnetic bead-based sandwich immunoassay was employed to capture target proteins.
  • DNA nanostructures were utilized as identifiable proxies, signaling the presence ('1') or absence ('0') of the captured target protein.
  • Solid-state nanopores were used as the sensing platform for detecting these DNA nanostructures.

Main Results:

  • The digital immunoassay scheme reliably quantified protein concentrations in complex biofluids.
  • The method demonstrated high sensitivity, achieving quantification down to the high femtomolar range for thyroid-stimulating hormone in human serum.
  • The approach successfully addressed specificity, sensitivity, and consistency challenges inherent in nanopore protein sensing.

Conclusions:

  • The developed digital immunoassay scheme offers a reliable method for protein quantification using solid-state nanopores.
  • This approach, utilizing DNA nanostructures as proxies, overcomes key challenges in nanopore-based protein sensing.
  • Further optimization holds promise for developing sensitive, high-dynamic-range diagnostic tools for point-of-care applications in precision medicine.