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Single molecule multiplexed nanopore protein screening in human serum using aptamer modified DNA carriers
Jasmine Y Y Sze1, Aleksandar P Ivanov2, Anthony E G Cass1
1Department of Chemistry, Imperial College London, Exhibition Road, London, SW7 2AZ, UK.
Nature Communications
|November 18, 2017
Summary
This study introduces a novel, low-cost nanopore sensing platform for simultaneous single-molecule protein detection in biological fluids. The method uses DNA carriers and aptamers for high selectivity, enabling sensitive diagnostics.
Area of Science:
- Biotechnology
- Nanotechnology
- Molecular Diagnostics
Background:
- Screening proteins at the single-molecule level in biological fluids is crucial for advancing diagnostics and therapeutics.
- Current methods often require expensive labels or extensive sample preparation, limiting their accessibility.
- Nanopore sensing combined with nucleic acid aptamer recognition offers a promising label-free, multiplexing approach.
Purpose of the Study:
- To develop a flexible, scalable, and low-cost detection platform for simultaneous multi-protein screening at the single-molecule level.
- To demonstrate the utility of this platform in complex biological matrices like human serum.
- To achieve ultra-low concentration detection of protein targets.
Main Methods:
- Utilizing a double-stranded DNA carrier backbone with grafted nucleic acid aptamer sequences.
- Employing nanopore sensing to detect unique ionic current signatures generated by protein-aptamer binding.
- Analyzing characteristic sub-peak current changes to differentiate protein sizes and enable target recognition.
Main Results:
- Successfully demonstrated simultaneous detection of multiple protein targets.
- Achieved accurate protein recognition through unique ionic current signatures.
- Differentiated individual protein sizes based on characteristic current changes.
- Enabled single-molecule screening in human serum at ultra-low protein concentrations.
Conclusions:
- The developed DNA-carrier-based nanopore sensing platform is a versatile, scalable, and cost-effective solution for multiplexed single-molecule protein detection.
- This approach significantly advances the potential for sensitive and selective diagnostics in complex biological samples.
- The method overcomes limitations of traditional techniques, paving the way for improved diagnostic and therapeutic strategies.
