Coupling method

Patent metadata

Field	Value
Publication number	US-12473595-B2
Application number	US-202519094566-A
Country	US
Kind code	B2
Filing date	Mar 28, 2025
Priority date	May 27, 2011
Publication date	Nov 18, 2025
Grant date	Nov 18, 2025

Abstract

The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.

First claim

The invention claimed is:

1 . A method for detecting polymers comprising modified nucleotides, the method comprising:
(a) providing a membrane in which is present a nanopore that provides a channel through the membrane; (b) contacting the membrane, in an ionic solution, with polymers comprising modified nucleotides, wherein following contact with the membrane the polymers are tethered to the membrane via a tethering group; (c) applying a potential difference across the membrane and detecting a first polymer using the nanopore, from among the polymers tethered to the membrane; and (d) detecting a second polymer, from among the polymers tethered to the membrane, using the same nanopore, wherein the second polymer is not the first polymer.

2 . The method of claim 1 , wherein the membrane is an amphiphilic layer.

3 . The method of claim 1 , wherein the membrane is a lipid bilayer.

4 . The method of claim 3 , wherein the lipid bilayer comprises a phosphatidylcholine (PC) or a phosphatidylethanolamine (PE) headgroup.

5 . The method of claim 1 , wherein the membrane is a block copolymer.

6 . The method of claim 5 , wherein the block copolymer is a triblock copolymer having outer hydrophilic layers and an inner hydrophobic core.

7 . The method of claim 1 , wherein the membrane comprises a mixture of lipids and copolymers.

8 . The method of claim 1 , wherein the nanopore is a protein nanopore.

9 . The method of claim 8 , wherein the protein nanopore is α-hemolysin (α-HL) nanopore.

10 . The method of claim 1 , wherein the polymers each comprise a tethering group.

11 . The method of claim 10 , wherein the tethering group is hydrophobic.

12 . The method of claim 1 , wherein polymers are each tethered to the membrane transiently.

13 . The method of claim 1 , wherein the polymers each comprises a leader sequence designed to preferentially thread into the nanopore.

14 . The method of claim 1 , wherein the polymers each further comprises polyethylene glycol (PEG).

15 . The method of claim 1 , wherein the detecting comprises measuring a current passing through the nanopore.

16 . The method of claim 1 , wherein the detecting comprises an electrical measurement.

17 . The method of claim 1 , wherein the detecting comprises measuring impedance.

18 . The method of claim 1 , wherein the detecting comprises measuring capacitance.

Assignees

• Oxford Nanopore Tech Plc

Inventors

• Clarke James Anthony
• White James
• Milton John
• Brown Clive Gavin

Classifications

• G01N27/44743

  Introducing samples · CPC title

• G01N33/48721

  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) · CPC title

• B82Y15/00

  Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors · CPC title

• C12Q1/68

  involving nucleic acids · CPC title

• G01N33/487

  of liquid biological material · CPC title