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Protein detection using tunable pores: resistive pulses and current rectification.

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Summary

This study compares two nanopore assays for detecting the cancer biomarker Vascular Endothelial Growth Factor (VEGF). Both methods, resistive pulses and current rectification, show comparable sensitivity for VEGF detection.

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Area of Science:

  • Nanotechnology
  • Biomarker Detection
  • Biosensing

Background:

  • Nanopore sensing offers a promising platform for sensitive biomolecule detection.
  • Quantifying cancer biomarkers like Vascular Endothelial Growth Factor (VEGF) is crucial for early diagnosis and treatment monitoring.
  • Existing nanopore assays require further optimization for sensitivity and ease of use.

Purpose of the Study:

  • To compare two distinct nanopore assay formats for quantifying the cancer biomarker VEGF.
  • To evaluate the sensitivity and applicability of resistive pulse and current rectification assays on a tunable pore platform.
  • To demonstrate the utility of Layer-by-Layer (LbL) assembly for aptamer immobilization in nanopore sensing.

Main Methods:

  • Developed and compared two nanopore assay strategies: resistive pulse measurements of nanoparticle translocation and current rectification analysis of functionalized pores.
  • Utilized aptamer-modified nanoparticles to measure electrophoretic mobility changes for VEGF detection (down to 18 pM).
  • Employed Layer-by-Layer (LbL) assembly to immobilize DNA aptamers onto conical nanopores, exploiting current rectification properties for VEGF detection (down to 5 pM).

Main Results:

  • The resistive pulse assay detected VEGF by monitoring aptamer-functionalized nanoparticle translocation speed, achieving a sensitivity of 18 pM.
  • The current rectification assay, utilizing LbL assembly of aptamers on nanopores, demonstrated pH and ionic strength-dependent currents and detected VEGF with a sensitivity of 5 pM.
  • Both assay formats exhibited comparable sensitivities for VEGF quantification, highlighting their potential in biomarker detection.

Conclusions:

  • Both resistive pulse and current rectification nanopore assays provide sensitive and comparable detection of the cancer biomarker VEGF.
  • Layer-by-Layer (LbL) assembly offers a facile method for aptamer immobilization, enhancing nanopore sensor specificity and sensitivity.
  • The tunable pore platform supports diverse assay formats, paving the way for advanced diagnostic tools.