Nanopore-Based Multiparametric Single-Molecule Protein Detection: From Structural Analysis to Clinical Translation

Area of Science:

• Biophysics
• Analytical Chemistry
• Biotechnology

Background:

• Single-molecule protein analysis is vital for understanding biological mechanisms and early disease detection.
• Nanopore sensing technology provides label-free, high-throughput, and direct electrical readout for protein analysis.

Purpose of the Study:

• This review summarizes recent advancements in nanopore-based protein analysis.
• It focuses on detecting both folded and unfolded proteins and their characteristics.

Main Methods:

• Analysis of ionic current blockade signals for protein size and shape.
• Monitoring protein conformational changes and unfolding processes.
• Detection of posttranslational modifications (PTMs) and sensitive quantitative analysis.

Main Results:

• Nanopore sensing enables detailed characterization of protein states, including unfolding and PTMs.
• Integration of machine learning enhances signal classification and analytical performance.
• Successful applications in complex biological samples like serum and urine have been demonstrated.

Conclusions:

• Nanopore technology is a powerful tool for protein analysis, with significant potential for clinical translation.
• Future directions include integrating nanopore arrays, optical readout, and machine learning to improve performance and accelerate clinical applications.