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Monitoring Protein Adsorption with Solid-state Nanopores

Monitoring Protein Adsorption with Solid-state Nanopores

Published on: December 2, 2011

High-bandwidth protein analysis using solid-state nanopores.

Joseph Larkin¹, Robert Y Henley¹, Murugappan Muthukumar²

¹Departments of Physics and Chemistry/Chemical Biology, Northeastern University, Boston, Massachusetts.

Biophysical Journal

|February 11, 2014

Summary

This summary is machine-generated.

High-bandwidth nanopore measurements enable precise analysis of small proteins. This study reveals confinement-driven interactions and provides accurate volumetric information, advancing protein detection capabilities.

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Area of Science:

Nanotechnology
Biophysics
Analytical Chemistry

Background:

Accurate analysis of small protein molecules is crucial for biological research.
Existing nanopore sensing methods face limitations in time resolution and detection efficiency for sub-30 kD proteins.

Purpose of the Study:

To develop high-bandwidth nanopore measurements for analyzing sub-30 kD protein molecules.
To investigate protein translocation dynamics, diffusion behavior, and pore interactions.
To determine protein volumetric information using nanopore sensing.

Main Methods:

Utilizing hafnium oxide and silicon nitride nanopores for high-bandwidth ion current measurements.
Applying a 1D diffusion-drift model to analyze translocation time distributions.
Employing a Kramers model to assess confinement-driven pore/protein interactions.

Main Results:

Achieved unprecedented time resolution (2.5 μs) and detection efficiency for sub-30 kD proteins.
Demonstrated that protein drift velocity scales linearly with voltage, indicating electrophoretic movement.
Observed significantly reduced protein diffusion constants (D) due to confinement, suggesting pore/protein interactions (9-12 kJ/mol).
Material-dependent reductions in mobility (μ) and diffusion (D) were noted.
Obtained accurate volumetric information for proteins, consistent with dynamic light scattering.
Successfully detected a protein-protein complex.

Conclusions:

High-bandwidth nanopore measurements offer a powerful tool for detailed analysis of small proteins.
Confinement within nanopores significantly influences protein diffusion and interactions.
Nanopore sensing provides reliable volumetric data and can detect complex biomolecular assemblies.