Monitoring protein adsorption with solid-state nanopores

David J. Niedzwiecki, Liviu Movileanu

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Solid-state nanopores have been used to perform measurements at the single-molecule level to examine the local structure and flexibility of nucleic acids, the unfolding of proteins, and binding affinity of different ligands. By coupling these nanopores to the resistive-pulse technique, such measurements can be done under a wide variety of conditions and without the need for labeling. In the resistive-pulse technique, an ionic salt solution is introduced on both sides of the nanopore. Therefore, ions are driven from one side of the chamber to the other by an applied transmembrane potential, resulting in a steady current. The partitioning of an analyte into the nanopore causes a well-defined deflection in this current, which can be analyzed to extract single-molecule information. Using this technique, the adsorption of single proteins to the nanopore walls can be monitored under a wide range of conditions. Protein adsorption is growing in importance, because as microfluidic devices shrink in size, the interaction of these systems with single proteins becomes a concern. This protocol describes a rapid assay for protein binding to nitride films, which can readily be extended to other thin films amenable to nanopore drilling, or to functionalized nitride surfaces. A variety of proteins may be explored under a wide range of solutions and denaturing conditions. Additionally, this protocol may be used to explore more basic problems using nanopore spectroscopy.

Original languageEnglish (US)
Article numbere3560
JournalJournal of Visualized Experiments
Issue number58
DOIs
StatePublished - Jan 1 2011

Keywords

  • Bioengineering
  • Issue 58
  • Nanopore spectroscopy
  • Protein adsorption
  • Resistive-pulse technique
  • S/TEM
  • Single-molecule biophysics
  • Solid-state nanopore

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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