Detection of translocation events using graphene-based nanopore assemblies

Patent metadata

Field	Value
Publication number	US-9921181-B2
Application number	US-201414454704-A
Country	US
Kind code	B2
Filing date	Aug 7, 2014
Priority date	Jun 26, 2014
Publication date	Mar 20, 2018
Grant date	Mar 20, 2018

Abstract

Translocation events are sensed using composite nanopore assemblies including nanopores formed in graphene sheets. Single molecule detection and characterization and multi-molecule characterization and identification are provided using such assemblies. Multiple electrodes associated with nanofluidic sensors facilitate detection of ionic current through a nanopore as well as tunneling currents. Current signals of individual molecules are estimated from the combination of an ionic current signal through the nanopore and tunneling current signals obtained at specific locations within the nanopore.

First claim

What is claimed is:

1. A method for detecting translocation events associated with a first target molecule while avoiding multiple binding events with respect to the first target molecule, comprising:
obtaining an assembly including a first graphene sheet bounded by first and second solid-state membranes and a nanopore extending through the graphene sheet and each of the solid-state membranes, the nanopore having an axis, the graphene sheet being positioned at a selected position with respect to the nanopore axis;
bonding one or more receptors selective to the first target molecule only to the first graphene sheet;
introducing an electrolyte solution to the nanopore;
applying an electric potential across the nanopore, and
detecting ionic current through the nanopore.

2. The method of claim 1 , further including the step of detecting a tunneling current within the graphene sheet.

3. The method of claim 1 , further including the step of bonding a plurality of receptors selective to the first target molecule to the graphene sheet.

4. The method of claim 1 , wherein only an edge of the graphene sheet is exposed to the nanopore, further including the step of oxidizing the edge of the graphene sheet prior to bonding the one or more receptors.

5. The method of claim 1 , wherein the graphene sheet includes a portion protruding into the nanopore.

6. A method for the simultaneous measurement of ionic current through a nanopore and multiple tunneling currents, comprising:
obtaining an assembly including a plurality of graphene sheets in alternating sequence with a plurality of solid state membranes and a nanopore extending through the graphene sheets and solid state membranes;
introducing an electrolyte solution to the nanopore;
applying an electric potential across the nanopore;
detecting ionic current through the nanopore, and
detecting a plurality of tunneling currents within the graphene sheets simultaneously with the step of detecting ionic current through the nanopore.

7. The method of claim 6 , further including the step of estimating individual current signals of individual molecules from a combination of the detected ionic current through the nanopore and the detected plurality of tunneling currents.

8. The method of claim 7 , wherein the step of estimating individual current signals of individual molecules further includes obtaining tunneling current events from the detected plurality of tunneling currents and analyzing the tunneling current events to obtain molecule translocation currents consistent with both the detected ionic current through the nanopore and the tunneling current events.

9. The method of claim 8 , further including the step of selecting a single set of molecule translocation currents based on least variability of at least one criterion.

10. The method of claim 8 , wherein the at least one criterion includes molecule speed through the nanopore, magnitude of the tunneling currents, or shape of the tunneling currents.

11. The method of claim 6 , further including the step of determining molecular velocity within the nanopore by monitoring the tunneling currents as a function of time.

12. The method of claim 6 , further including the step of applying an electric potential to at least one of the graphene layers.

13. The method of claim 6 , further including the step of detecting changes in the tunneling currents and using the detected changes in tunneling currents to differentiate at least one of molecular conformations or molecular orientations within the nanopore.

14. The method of claim 6 , wherein one or more of the graphene sheets is comprised of a plurality of discrete graphene strips, each of the discrete graphene strips including at least one nanopore extending therethrough, each of the graphene sheets and solid state membranes including a plurality of nanopores, further including the steps of introducing the electrolyte solution to the plurality of nanopores, detecting the ionic current through the plurality of nanopores, and detecting a plurality of tunneling currents within the discrete graphene strips simultaneously with the step of detecting the ionic current through the plurality of nanopores.

15. A system comprising:
an assembly including a plurality of graphene layers in alternating sequence with a plurality of solid state membranes;
a plurality of nanopores extending through the graphene layers and the solid state membranes, and
a plurality of the graphene layers being electrically connected to one or more detectors configured for detecting tunneling currents within the graphene layers associated with charged molecules within the nanopores.

16. The system of claim 15 , wherein each graphene layer is comprised of a plurality of discrete graphene strips, each of the discrete graphene strips including a plurality of the nanopores extending therethrough, a plurality of the graphene strips being electrically connected to the one or more detectors.

17. The system of claim 16 , wherein the graphene strips comprising a first of the graphene layers have longitudinal axes that extend in a first direction and the graphene strips comprising a second of the graphene layers have longitudinal axes that extend in a second direction intersecting the first direction, one or more of the graphene strips comprising the first graphene layer overlapping one or more of the graphene strips comprising the second graphene layer.

18. The system of claim 16 , wherein one or more of the graphene strips comprising a first of the graphene layers overlap at least two of the graphene strips comprising a second of the graphene layers.

19. The system of claim 16 , further including electrolyte within the nanopores and means for detecting ionic current through the nanopores.

Assignees

• IBM

Inventors

• Baldauf Julia S
• Downton Matthew
• Gunn Natalie
• Harrer Stefan
• Kannam Sridhar
• Schieber Christine
• Wagner John M

Classifications

• G01N33/48721 Primary

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title

• G01N27/3276

  being a hybridisation with immobilised receptors (using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68) · CPC title

• C12Q1/6827 Primary

  for detection of mutation or polymorphism · CPC title

• G01N27/44791 Primary

  Microapparatus (sample containers with integrated microfluidic structures B01L3/5027) · CPC title

• C12Q2565/631

  being a biochannel or pore · CPC title