A5082906507- Nanopore and Nanochannel Transport Studies
- Microfluidic and Capillary Electrophoresis Applications
- Ion-surface interactions and analysis
- Microfluidic and Bio-sensing Technologies
- Electrostatics and Colloid Interactions
- Lipid Membrane Structure and Behavior
- Membrane-based Ion Separation Techniques
- Mass Spectrometry Techniques and Applications
- Erythrocyte Function and Pathophysiology
- Geophysical and Geoelectrical Methods
- Advanced biosensing and bioanalysis techniques
University of Groningen
2020-2023
VIB-KU Leuven Center for Microbiology
2023
KU Leuven
2022
Biotechnology Institute
2020-2021
Abstract Nanopores are single-molecule sensors used in nucleic acid analysis, whereas their applicability towards full protein identification has yet to be demonstrated. Here, we show that an engineered Fragaceatoxin C nanopore is capable of identifying individual proteins by measuring peptide spectra produced from hydrolyzed proteins. Using model proteins, the resulting experiments and mass spectrometry share similar profiles, hence allowing fingerprinting. The intensity peaks provides...
Although nanopores can be used for single-molecule sequencing of nucleic acids using low-cost portable devices, the characterization proteins and their modifications has yet to established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC recognized. However, high ionic strengths (i.e., 3 M LiCl) low pH 3) together with a nanopore phenylalanine at its constriction allows recognition peptides, distinguish between mono- diglycosylated peptides. Using...
The detection of analytes and the sequencing DNA using biological nanopores have seen major advances over recent years. analysis proteins peptides with nanopores, however, is complicated by complex physicochemical structure polypeptides lack understanding mechanism capture recognition nanopores. In this work, we show that introducing aromatic amino acids at precise positions within lumen α-helical fragaceatoxin C (FraC) increased frequency largely improved discrimination among similar size....
An extended PNP-NS framework for accurately simulating the nanofluidic properties of nanopores, yielding an in-depth description Cytolysin A (ClyA) and paving road towards rational engineering biological nanopores.
The ability to measure the concentration of metabolites in biological samples is important, both clinic and for home diagnostics. Here we present a nanopore-based biosensor automated data analysis quantification thiamine urine less than minute, without need recalibration. For this use Cytolysin A nanopore equip it with an engineered periplasmic binding protein (TbpA). To allow fast measurements tuned affinity TbpA by redesigning π-π stacking interactions between thiazole group TbpA. This...
Mass spectrometry (MS) is widely used in proteomic analysis but cannot differentiate between molecules with the same mass-to-charge ratio. Nanopore technology might provide an alternative method for rapid and cost-effective sequencing of proteins. In this study, we demonstrate that nanopore currents can distinguish diastereomeric enantiomeric differences l- d-peptides, not observed by conventional MS analysis, down to individual d-amino acids small opioid peptides. Molecular dynamics...
The identification of proteins at the single-molecule level would open exciting new venues in biological research and disease diagnostics. Previously, we proposed a nanopore-based method for protein called chop-n-drop fingerprinting, which fragmentation pattern induced measured by proteasome-nanopore construct is used to identify single proteins. In simulation study presented here, show that 97.1% human proteome constituents are uniquely identified under close ideal measuring circumstances,...
Single-molecule nanopore electrophysiology is an emerging technique for the detection of analytes in aqueous solutions with high sensitivity. These detectors have proven applicable enzyme-assisted sequencing oligonucleotides. There has recently been increased interest use nanopores fingerprinting peptides and proteins, referred to as single-molecule spectrometry. However, analysis resulting traces remains complicated due fast unassisted translocation such analytes, usually order micro-...
Novel technologies for high precision and temporal resolution of cellular metabolites.
Abstract In recent years, the protein nanopore cytolysin A (ClyA) has become a valuable tool for detection, characterization and quantification of biomarkers, proteins nucleic acids at single-molecule level. Despite this extensive experimental utilization, comprehensive computational study ion water transport through ClyA is currently lacking. Such yields wealth information on electrolytic conditions inside pore scale electrophoretic forces that drive molecular transport. To end we have...
Abstract The ability to measure the concentration of metabolites in biological samples is important, both clinic and for home diagnostics. Here we present a nanopore‐based biosensor automated data analysis quantification thiamine urine less than minute, without need recalibration. For this use Cytolysin A nanopore equip it with an engineered periplasmic binding protein (TbpA). To allow fast measurements tuned affinity TbpA by redesigning π‐π stacking interactions between thiazole group TbpA....