Systems and methods for forming a nanopore in a lipid bilayer

• US 9,605,307 B2
• Filed: 02/08/2010
• Issued: 03/28/2017
• Est. Priority Date: 02/08/2010
• Status: Active Grant

• Current Assignee: Roche Sequencing Solutions, Inc. (Roche Holding AG)
• Original Assignee: Genia Technologies, Inc. (Roche Holding AG)
• Inventors: Chen, Roger, Davis, Randy
• Primary Examiner(s): RUFO, LOUIS J
• Application Number:
  US12/658,591
• Publication Number:
  US 20110193249A1
• Time in Patent Office: 2,605 Days
• Field of Search: 204/450
• US Class Current: 1/1
• CPC Class Codes: B29C 67/20 for porous or cellular articles, e.g. of foam plastics, coarse-pored chemical aspects of working up macro-molecular substances to porous or cellular articles C08J9/00
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: 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
• CPC Class Codes: Y10S 977/797 Lipid particle
• CPC Class Codes: Y10S 977/90 having step or means utilizing mechanical or thermal property, e.g. pressure, heat
• CPC Class Codes: Y10S 977/942 including protein logic element

Abstract

Techniques for forming a nanopore in a lipid bilayer are described herein. In one example, an agitation stimulus level such as an electrical agitation stimulus is applied to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer. In some embodiments, a change in an electrical property of the lipid bilayer resulting from the formation of the nanopore in the lipid bilayer is detected, and a nanopore has formed in the lipid bilayer is determined based on the detected change in the lipid bilayer electrical property.

Claims

1. A system for forming a nanopore in a lipid bilayer characterized by a resistance including:

a substantially planar solid surface including a lipid bilayer compatible planar surface portion on which the lipid bilayer forms;

a source that is coupled to the substantially planar solid surface;

a sensing circuit coupled to the source; and

a processor;

wherein the processor is programmed to;

determine that a lipid bilayer is formed on the lipid bilayer compatible planar surface portion based on a first measurement from the sensing circuit;

in response to the determination that the lipid bilayer is formed and after nanopore forming molecules are deposited on the lipid bilayer, control the source to apply a first agitation electrical stimulus level to the lipid bilayer for up to one second, causing a disruption in the lipid bilayer that tends to facilitate the formation of nanopores in the lipid bilayer;

control the source to apply a second negative electrical stimulus level immediately following the first agitation electrical stimulus level to the lipid bilayer, causing reverse oxidation of at least a portion of the system; and

control the source to apply a third electrical stimulus level following the second negative electrical stimulus level to the lipid bilayer and determine that a nanopore has been formed based on a second measurement received from the sensing circuit sensing in response to the third electrical stimulus level, wherein an absolute magnitude of the first agitation electrical stimulus level is at least two times greater than an absolute magnitude of the third electrical stimulus level.

2. The system of claim 1, further comprising an additional source, and wherein the processor is programmed to:

in response to the determination that the lipid bilayer is formed and after nanopore forming molecules are deposited on the lipid bilayer, control the additional source to apply an additional agitation stimulus to the lipid bilayer, causing a disruption in the lipid bilayer that tends to facilitate the formation of nanopores in the lipid bilayer, wherein the additional agitation stimulus is selected from the group consisting of;

a mechanical stimulus, a sound stimulus, a chemical light stimulus, and a thermal stimulus.

3. The system of claim 1, wherein the first agitation electrical stimulus level tends to facilitate the insertion of an α

-hemolysin nanopore in the lipid bilayer.

4. The system of claim 1, wherein:

the sensing circuit is configured to detect a change in the electrical property of the lipid bilayer resulting from the formation of a nanopore in the lipid bilayer; and

wherein the processor is programmed to determine that a nanopore has formed in the lipid bilayer based on the detected change in the lipid bilayer electrical property.

5. The system of claim 4, wherein detecting a change in the lipid bilayer electrical property comprises detecting a change in a resistance of the lipid bilayer.

6. The system of claim 4, wherein determining that a nanopore is formed includes determining a number of nanopores formed based on a size of change in the lipid bilayer electrical property.

7. The system of claim 6, the processor is further programmed to control the source to apply an erasing electrical stimulus to erase the lipid bilayer when it is determined that more than one nanopore is formed in the lipid bilayer.

8. The system of claim 6, the processor is further programmed to control the source to apply another agitation electrical stimulus level to a lipid bilayer wherein the agitation electrical stimulus level tends to facilitate the formation of nanopores in the lipid bilayer when it is determined that no nanopore is formed in the lipid bilayer.

9. The system of claim 4, wherein detecting a change in the electrical property of the lipid bilayer resulting from the formation of a nanopore in the lipid bilayer includes detecting a decrease in resistance of the lipid bilayer.

10. The system of claim 1, wherein the substantially planar solid surface comprises a substantially planar hydrophilic solid surface.

11. The system of claim 1, wherein the substantially planar solid surface comprises a substantially planar electrode surface.

12. The system of claim 1, wherein the system is automated.

13. The system of claim 1, wherein the nanopore is one of a plurality of nanopores in a nanopore array.

14. The system of claim 13, wherein each of the plurality of nanopores is individually addressable.

15. The system of claim 13, wherein each of the plurality of nanopores is individually controllable.