Graphene supported artificial membranes and uses thereof

Patent metadata

Field	Value
Publication number	US-9797863-B2
Application number	US-201414478820-A
Country	US
Kind code	B2
Filing date	Sep 5, 2014
Priority date	Jun 8, 2010
Publication date	Oct 24, 2017
Grant date	Oct 24, 2017

Abstract

The invention features the use of graphene, a one atom thick planar sheet of bonded carbon atoms, in the formation of artificial lipid membranes. The invention also features the use of these membranes to detect the properties of polymers (e.g., the sequence of a nucleic acid) and identify transmembrane protein-interacting compounds.

First claim

What is claimed is:

1. A device comprising an artificial membrane spanning one or more apertures on graphene and at least one transmembrane pore disposed in the artificial membrane in one of said one or more apertures, wherein the membrane is a bilayer within the one or more apertures and two monolayers interleaved by the graphene at the peripheral edges of the one or more apertures.

2. The device of claim 1 , wherein said transmembrane pore comprises a transmembrane protein.

3. The device of claim 2 , wherein said transmembrane protein is selected from the group consisting of ion channels, multidrug resistance pumps, cytokine receptors, receptors from the immunoglobin superfamily, receptors from the tumor necrosis factor receptor family, chemokine receptors, receptor tyrosine kinases, and TGF beta receptors.

4. The device of claim 1 , wherein at least one of said pores is α-haemolysin.

5. The device of claim 1 , further comprising a frame supporting said graphene.

6. The device of claim 5 , further comprising first and second electrodes disposed on opposite sides of said graphene.

7. The device of claim 5 , wherein said frame comprises a ceramic or silicon nitride.

8. The device of claim 1 , wherein said device comprises a plurality of apertures arranged in an array.

9. The device of claim 8 , further comprising a frame positioned on one face of the graphene, wherein said frame forms separate compartments for housing liquid for each of said apertures.

10. The device of claim 9 , wherein said frame is non-conductive.

11. The device of claim 9 , wherein said frame comprises a ceramic or silicon nitride.

12. The device of claim 9 , wherein each of said compartments further comprises an electrode, and wherein said device further comprises at least one electrode on the side of said graphene opposite said compartment.

13. The device of claim 1 wherein the at least one of said pores is OmpF, OmpC or MspA.

14. The device of claim 13 wherein the artificial membrane is a lipid bilayer.

15. The device of claim 1 , wherein at most one pore is disposed in each of said one or more apertures.

16. The device of claim 1 , further comprising a molecular motor, wherein said motor is adjacent to the transmembrane pore and is capable of moving a polymer with respect to the transmembrane pore.

17. The device of claim 16 , wherein the molecular motor comprises a DNA polymerase, a RNA polymerase, a ribosome, an exonuclease, or a helicase and said polymer is a polynucleotide.

18. The device of claim 17 , wherein the DNA polymerase is selected from E. coli DNA polymerase I, E. coli DNA polymerase I Large Fragment (Klenow fragment), phage T7 DNA polymerase, Phi-29 DNA polymerase, Thermus aquaticus (Taq) DNA polymerase, Thermus flavus (Tfl) DNA polymerase, Thermus Thermophilus (Tth) DNA polymerase, Thermococcus litoralis (Tli) DNA polymerase, Pyrococcus furiosus (Pfu) DNA polymerase, Bacillus stearothermophilus (Bst) DNA polymerase, AMV reverse transcriptase, MMLV reverse transcriptase, and HIV-1 reverse transcriptase.

19. The device of claim 17 , wherein the RNA polymerase is selected from T7 RNA polymerase, T3 RNA polymerase, SP6 RNA polymerase, and E. coli RNA polymerase.

20. The device of claim 17 , wherein the exonuclease is selected from exonuclease Lambda, T7 Exonuclease, Exo III, RecJ 1 Exonuclease, Exo I, and Exo T.

21. The device of claim 17 , wherein the helicase is selected from E - coli bacteriophage T7 gp4 and T4 gp41 gene proteins, E. coli protein DnaB, E. coli protein RuvB, and E. coli protein rho.

22. A method of identifying a membrane molecule-interacting compound comprising providing a device comprising an artificial membrane spanning one or more apertures on graphene and said membrane molecule, contacting the membrane molecule with a compound, and measuring at least one activity of said membrane molecule in the presence of said compound, whereby a change in activity in the presence of said compound indicates that said compound interacts with said membrane molecule, wherein said membrane molecule is a transmembrane pore disposed in the artificial membrane in one of said one or more apertures, wherein measuring at least one activity of said membrane molecule in the presence of said compound comprises measuring electric current through or across the one or more apertures, and wherein the membrane is a bilayer within the one or more apertures and two monolayers interleaved by the graphene at the peripheral edges of the one or more apertures.

23. The method of claim 22 , wherein membrane molecule-interacting compound is a polymer, and the device further comprises a molecular motor, wherein said motor is adjacent to the transmembrane pore and is capable of moving the polymer with respect to the transmembrane pore.

24. The method of claim 22 , wherein the transmembrane pore comprises a transmembrane protein.

Assignees

• Harvard College

Inventors

• Garaj Slaven
• Branton Daniel

Classifications

• G01N27/447

  using electrophoresis · CPC title

• Y10S977/734

  Fullerenes, i.e. graphene-based structures, such as nanohorns, nanococoons, nanoscrolls or fullerene-like structures, e.g. WS2 or MoS2 chalcogenide nanotubes, planar C3N4, etc. · CPC title

• G01N27/44791 Primary

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

• G01N27/453

  Cells therefor · CPC title

• 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