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Single-Molecule Protein Profiling Using Nanopores and Dimeric Aptamer-Modified DNA Carriers.

Aleksandar Ivanov1,2, Xiaoyi Wang1, Yaxian Liu1,3

  • 1Department of Chemistry, Imperial College London, Molecular Sciences Research Hub, White City Campus, 82 Wood Lane, London, W12 0BZ, UK.

Angewandte Chemie (International Ed. in English)
|June 29, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a novel nanopore sensor strategy for highly sensitive, single-molecule protein detection. The method uses aptamer-protein-aptamer sandwiches to achieve sub-picomolar limits for detecting biomarkers like VEGF and PDGF.

Keywords:
AptamersDiagnosticsMolecular carriersNanoporesSingle‐molecule protein detection

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Biomarker Discovery

Background:

  • Single-molecule protein profiling offers high sensitivity for rare biomarker detection, crucial for early disease diagnosis.
  • Nanopore sensors provide sensitive, high-throughput protein detection in solution, suitable for clinical use.

Purpose of the Study:

  • To develop an enhanced nanopore-based detection strategy for improved protein profiling sensitivity.
  • To enable selective detection and characterization of specific proteins and protein interactions.

Main Methods:

  • Utilized an aptamer-protein-aptamer sandwich structure with DNA carriers for enhanced protein transport and detection.
  • Analyzed charge-based dimerization of DNA carriers and characteristic subpeaks in nanopore events for protein identification.
  • Correlated subpeak height to protein size for quantitative analysis.

Main Results:

  • Achieved selective detection of dimeric proteins, including vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), at sub-picomolar limits.
  • Demonstrated adaptability for proteins with two aptamer-binding sites, exemplified by thrombin detection.
  • Enabled real-time monitoring of ligand-induced receptor dimerization in complex biological fluids.

Conclusions:

  • The developed nanopore strategy significantly enhances single-molecule protein detection sensitivity and selectivity.
  • This method provides a powerful tool for potential single-molecule-based diagnostics and monitoring of protein interactions.
  • The approach is versatile and adaptable for a wide range of protein targets and clinical applications.