Multiplexed continuous biosensing by single-molecule encoded nanoswitches

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Single-molecule techniques have become impactful in bioanalytical sciences, though the advantages for continuous biosensing are yet to be discovered. Here we present a multiplexed, continuous biosensing method, enabled by an analyte-sensitive, single-molecular nanoswitch with a particle as a reporter. The nanoswitch opens and closes under the influence of single target molecules. This reversible switching yields binary transitions between two highly reproducible states, enabling reliable quantification of the single-molecule kinetics. The multiplexing functionality is encoded per particle via the dissociation characteristics of the nanoswitch, while the target concentration is revealed by the association characteristics. We demonstrate by experiments and simulations the multiplexed, continuous monitoring of oligonucleotide targets, at picomolar concentrations in buffer and in filtered human blood plasma.
Original languageEnglish
Pages (from-to)2296-2302
Number of pages7
JournalNano Letters
Issue number4
Early online date12 Mar 2020
Publication statusPublished - 8 Apr 2020


  • continuous biosensing
  • kinetic identification
  • multiplexing
  • single-molecule biosensing


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