Concentrating a target molecule for sensing by a nanopore

Patent metadata

Field	Value
Publication number	US-11920184-B2
Application number	US-202017071778-A
Country	US
Kind code	B2
Filing date	Oct 15, 2020
Priority date	Nov 9, 2012
Publication date	Mar 5, 2024
Grant date	Mar 5, 2024

Abstract

Methods and related products are disclosed that improve the probability of interaction between a target molecule and a nanopore by capturing the target molecule on a surface comprising the nanopore. The captured target molecule, the nanopore, or both, are able to move relative to each other along the surface. When the leader of the target molecule is in proximity with the nanopore, interaction of the target portion of the target molecule with the nanopore occurs, thereby permitting sensing of the target portion. Confining the target molecule and nanopore in this manner leads to significantly enhanced interaction with the nanopore.

First claim

The invention claimed is:

1. A method for concentrating a target molecule for nanopore sensing, comprising: capturing the target molecule on a surface, wherein the surface comprises a nanopore and a hydrophobic domain, wherein the target molecule comprises a target portion, a hydrophobic capture element and a leader for interaction with the nanopore, wherein the target portion is an Xpandomer, wherein the Xpandomer is a measurable polymer comprising high signal-to-noise reporters that are separated by approximately 10 nm, wherein the hydrophobic capture element is an aliphatic hydrocarbon, wherein the aliphatic hydrocarbon comprises 5 repeats of a 12 carbon monomer, and wherein the leader is a hydrophobic polymer, wherein the hydrophobic polymer comprises 25 repeats of an ethyl phosphodiester, wherein the hydrophobic capture element is associated with, and capable of movement along, the hydrophobic domain of the surface to bring the leader in proximity with the nanopore; and sensing at least the target portion upon interaction with the nanopore.

2. The method of claim 1 , wherein the step of capturing the target molecule on the surface comprises contacting the surface with the target molecule, wherein the target molecule comprises, prior to the contacting step, the target portion, the hydrophobic capture element and the leader.

3. The method of claim 1 , wherein the step of capturing the target molecule on the surface comprises linking the hydrophobic capture element associated with the surface to the target portion and leader, thereby capturing the target molecule on the surface. nanopore.

4. The method of claim 1 , wherein the nanopore is a biological nanopore.

5. The method of claim 1 , wherein the surface is a lipid bilayer.

6. The method of claim 1 , wherein the surface is a solid-state or synthetic membrane.

7. The method of claim 1 , wherein the target portion comprises a molecular bar code.

8. The method of claim 1 , wherein the target molecule comprises two or more hydrophobic capture elements.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Kokoris Mark Stamatios
• Mcruer Robert N

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• G01N33/54373

  involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings · CPC title

• C12Q2563/131

  the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin · CPC title

• C12Q2565/531

  characterised by the capture moiety being a protein for target oligonucleotides · CPC title