# US 2015/0101931 A1 · Graphene supported artificial membranes and uses thereof

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- Number: US 2015/0101931 A1
- Document ID: US20150101931A1
- Status: Published application
- Kind: A1
- Application number: 14/478,820
- Filed: 2014-09-05
- Published: 2015-04-16
- Earliest claimed: 2010-06-08
- Assignees: President and Fellows of Harvard College
- Applicants: President and Fellows of Harvard College
- Inventors: Slaven GARAJ; Daniel BRANTON
- Web page: https://patentgrep.com/patent/US20150101931A1

## CPC

- G01N 27/44791: {Microapparatus (sample containers with integrated microfluidic structures B01L3/5027)}
- G01N 27/447: using electrophoresis
- G01N 27/453: Cells therefor
- G01N 33/48721: {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)}
- Y10S 977/734: 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.
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## 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.

## Claims

### Claim 1

A device comprising an artificial membrane located within one or more apertures on graphene.

### Claim 2 (depends on 1)

The device of claim 1, wherein said artificial membrane further comprises a protein.

### Claim 3 (depends on 2)

The device of claim 2, wherein said protein is a transmembrane protein.

### Claim 4 (depends on 3)

The device of claim 3, 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.

### Claim 5 (depends on 1)

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

### Claim 6 (depends on 5)

The device of claim 5, further comprising a molecular motor, wherein said motor is adjacent to at least one of said one or more pores and is capable of moving a polymer with respect to the at least one of said one or more pores.

### Claim 7 (depends on 6)

The device of claim 6, wherein the molecular motor comprises a DNA polymerase, a RNA polymerase, a ribosome, an exonuclease, or a helicase and said polymer is a polynucleotide.

### Claim 8 (depends on 7)

The device of claim 7, 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.

### Claim 9 (depends on 7)

The device of claim 7, wherein the RNA polymerase is selected from T7 RNA polymerase, T3 RNA polymerase, SP6 RNA polymerase, and E. coli RNA polymerase.

### Claim 10 (depends on 7)

The device of claim 7, wherein the exonuclease is selected from exonuclease Lambda, T7 Exonuclease, Exo III, RecJ1 Exonuclease, Exo I, and Exo T.

### Claim 11 (depends on 7)

The device of claim 7, 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.

### Claim 12 (depends on 5)

The device of claim 5, wherein at least one of said pores is α-haemolysin.

### Claim 13 (depends on 1)

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

### Claim 14 (depends on 13)

The device of claim 13, further comprising first and second electrodes disposed on opposite sides of said graphene.

### Claim 15 (depends on 13)

The device of claim 13, wherein said frame comprises a ceramic or silicon nitride.

### Claim 16 (depends on 1)

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

### Claim 17 (depends on 16)

The device of claim 16, 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.

### Claim 18 (depends on 17)

The device of claim 17, wherein said frame is non-conductive.

### Claim 19 (depends on 17)

The device of claim 17, wherein said frame comprises a ceramic or silicon nitride.

### Claim 20 (depends on 17)

The device of claim 17, 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.

### Claim 21 (depends on 5)

A method of analyzing a target polymer comprising introducing the target polymer to the device of claim 5, allowing the target polymer to move with respect to the pore to produce a signal, and monitoring the signal corresponding to the movement of the target polymer with respect to the pore, thereby analyzing the target polymer.

### Claim 22 (depends on 21)

The method of claim 21, wherein the signal monitoring comprises measuring a monomer-dependent characteristic of the target polymer while the target polymer moves with respect to the pore.

### Claim 23 (depends on 22)

The method of claim 22, wherein the monomer dependent property is the identity of a monomer or the number of monomers in the polymer.

### Claim 24 (depends on 20)

The method of claim 20, further comprising altering the rate of movement of the polymer before, during, or after the signal monitoring.

### Claim 25 (depends on 1)

A method of identifying a membrane molecule-interacting compound comprising providing the device of claim 1 comprising 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.

### Claim 26 (depends on 25)

The method of claim 25, wherein an electric field is applied and said measuring at least one activity of said membrane molecule comprises measuring the electrical resistance of said membrane molecule.

### Claim 27 (depends on 25)

The method of claim 25, wherein said membrane molecule is a transmembrane protein.