Protein detection by nanopores equipped with aptamers

Dvir Rotem, Lakmal Jayasinghe, Maria Salichou, Hagan Bayley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

265 Scopus citations

Abstract

Protein nanopores have been used as stochastic sensors for the detection of analytes that range from small molecules to proteins. In this approach, individual analyte molecules modulate the ionic current flowing through a single nanopore. Here, a new type of stochastic sensor based on an αHL pore modified with an aptamer is described. The aptamer is bound to the pore by hybridization to an oligonucleotide that is attached covalently through a disulfide bond to a single cysteine residue near a mouth of the pore. We show that the binding of thrombin to a 15-mer DNA aptamer, which forms a cation-stabilized quadruplex, alters the ionic current through the pore. The approach allows the quantification of nanomolar concentrations of thrombin, and provides association and dissociation rate constants and equilibrium dissociation constants for thrombin•aptamer interactions. Aptamer-based nanopores have the potential to be integrated into arrays for the parallel detection of multiple analytes.

Original languageAmerican English
Pages (from-to)2781-2787
Number of pages7
JournalJournal of the American Chemical Society
Volume134
Issue number5
DOIs
StatePublished - 8 Feb 2012

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