Method of characterising a target polypeptide using a nanopore

Patent metadata

Field	Value
Publication number	US-2026072009-A1
Application number	US-202519396980-A
Country	US
Kind code	A1
Filing date	Nov 21, 2025
Priority date	Dec 2, 2019
Publication date	Mar 12, 2026
Grant date	—

Abstract

Provided herein are methods of characterising a target polypeptide as it moves with respect to a nanopore. Also provided are related kits, systems and apparatuses for carrying out such methods.

First claim

What is claimed:

1 . A method of controlling movement of a target polypeptide through a transmembrane protein pore, the method comprising:
(i) holding at least a portion of the target polypeptide in a linearized form in the transmembrane protein pore under electro-osmotic force conditions; (ii) controlling the movement of the target polypeptide through the transmembrane protein pore under an applied electrical potential; and (iii) taking one or more electrical measurements as the target polypeptide moves through the transmembrane protein pore.

2 . The method of claim 1 , wherein the electro-osmotic force conditions comprise an unbalanced flow of cations and anions through the transmembrane protein pore under the applied electrical potential.

3 . The method of claim 1 , wherein the transmembrane protein pore comprises one or more amino acid additions, substitutions, or deletions relative to an unmodified transmembrane protein pore.

4 . The method of claim 1 , wherein the electro-osmotic force conditions comprise asymmetric salt conditions across the transmembrane pore.

5 . The method of claim 1 , wherein the method is carried out in the presence of guanidine HCl.

6 . The method of claim 1 , wherein the target polypeptide is conjugated to a leader comprising a charged polymer.

7 . The method of claim 1 , wherein the target polypeptide is conjugated to a leader comprising a polynucleotide.

8 . The method of claim 1 , wherein the transmembrane protein pore is derived from MspA, MspB, MspC, MspD, α-hemolysin, lysenin, CsgG, OmpF, OmpG, NalP, ClyA, Sp1, or FraC.

9 . The method of claim 1 , wherein the transmembrane protein pore is derived from CsgG.

10 . The method of claim 1 , wherein the transmembrane protein pore is derived from MspA.

11 . The method of claim 1 , wherein taking the one or more electrical measurements comprises measuring an ion current flow through the transmembrane protein pore.

Assignees

• Oxford Nanopore Tech Plc

Inventors

• Heron Andrew John
• Graham James Edward
• Strycharska Melania Slawa

Classifications

• G01N2458/00

  Labels used in chemical analysis of biological material · CPC title

• G01N33/6818 Primary

  Sequencing of polypeptides · CPC title

• C12N2310/3517

  Marker; Tag · CPC title

• C12N15/11

  DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} · 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