Systems and methods for forming a nanopore in a lipid bilayer

• US 20110193249A1
• Filed: 02/08/2010
• Published: 08/11/2011
• 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
• Granted Patent: US 9,605,307 B2
• Time in Patent Office: Days
• US Class Current: 264/4
• 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 method of forming a nanopore in a lipid bilayer including:

applying an agitation stimulus level to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer.

2. The method of claim 1, wherein the agitation stimulus is selected from the group consisting of:

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

3. The method of claim 1 wherein the agitation stimulus comprises a voltage (V) level.

4. The method of claim 1, wherein the agitation stimulus level tends to facilitate the insertion of α

-hemolysin nanopore in the lipid bilayer.

5. The method of claim 1, further comprising:

detecting a change in an electrical property of the lipid bilayer resulting from the formation of a nanopore in the lipid bilayer;

determining that a nanopore has formed in the lipid bilayer based on the detected change in the lipid bilayer electrical property.

6. The method of claim 5, wherein one or more steps are automated.

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

8. The method of claim 5, determining that a nanopore is formed includes determining the number of nanopores formed based on a size of change in the bilayer electrical property.

9. The method of claim 8, further including applying an erasing electrical stimulus to erase the lipid bilayer when it is determined that more than one nanpore is formed in the lipid bilayer.

10. The method of claim 8, further including applying 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.

11. The method of claim 1, wherein detecting a change in the electrical property of the bilayer resulting from the formation of a nanopore in the lipid bilayer includes detecting a decrease in resistance of the bilayer.

12. The method of claim 1, wherein the lipid bilayer is formed on a substantially planar lipid bilayer compatible solid surface.

13. The method of claim 1, wherein the lipid bilayer is formed on a substantially planar hydrophilic solid surface.

14. The method of claim 1, wherein the lipid bilayer is formed on a substantially planar electrode surface.

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

a source configured to apply an agitation stimulus level to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer.

16. The system of claim 15, wherein the agitation stimulus is selected from the group consisting of:

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

17. The system of claim 15, wherein the agitation stimulus comprises an agitation electrical stimulus and the source comprises a voltage source configured to apply an agitation electrical stimulus level to the lipid bilayer wherein the agitation electrical stimulus level tends to facilitate the formation of nanopores in the lipid bilayer.

18. The system of claim 15, wherein the agitation stimulus level tends to facilitate the insertion of α

-hemolysin nanopore in the lipid bilayer.

19. The system of claim 15, further comprising:

a sensing circuit 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

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

20. The system of claim 19, wherein detecting a change in the lipid bilayer electrical property comprises detecting a change in a resistance of the lipid bilayer.

21. The system of claim 19, determining that a nanopore is formed includes determining the number of nanopores formed based on a size of change in the bilayer electrical property.

22. The system of claim 21, the variable voltage source is further configured 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.

23. The system of claim 21, the variable voltage source is further configured 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.

24. The system of claim 19, wherein detecting a change in the electrical property of the bilayer resulting from the formation of a nanopore in the lipid bilayer includes detecting a decrease in resistance of the bilayer.

25. The system of claim 15, wherein the lipid bilayer is formed on a substantially planar lipid bilayer compatible solid surface.

26. The system of claim 15, wherein the lipid bilayer is formed on a substantially planar hydrophilic solid surface.

27. The system of claim 15, wherein the lipid bilayer is formed on a substantially planar electrode surface.

28. The system of claim 15, wherein the system is automated.

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

means for applying an agitation stimulus level to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer.

30. The system of claim 29, wherein the agitation stimulus is selected from the group consisting of:

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

31. The system of claim 29, wherein the agitation stimulus comprises an electrical agitation stimulus.

32. The system of claim 29, wherein the agitation stimulus level tends to facilitate the insertion of α

-hemolysin nanopore in the lipid bilayer.

33. The system of claim 29, further comprising:

means for detecting a change in the electrical property of the lipid bilayer resulting from the formation of a nanopore in the lipid bilayer; and

means for determining that a nanopore has formed in the lipid bilayer based on the detected change in the lipid bilayer electrical property.

34. The system of claim 33, wherein detecting a change in the lipid bilayer electrical property comprises detecting a change in a resistance of the lipid bilayer.

35. The system of claim 33, determining that a nanopore is formed includes determining the number of nanopores formed based on a size of change in the bilayer electrical property.

36. The system of claim 35, further including means for applying an erasing electrical stimulus to erase the lipid bilayer when it is determined that more than one nanopore is formed in the lipid bilayer.

37. The system of claim 35, further including means for applying 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.

38. The system of claim 33, wherein detecting a change in the electrical property of the bilayer resulting from the formation of a nanopore in the lipid bilayer includes detecting a decrease in resistance of the bilayer.

39. The system of claim 29, wherein the lipid bilayer is formed on a substantially planar lipid bilayer compatible solid surface.

40. The system of claim 29, wherein the lipid bilayer is formed on a substantially planar hydrophilic solid surface.

41. The system of claim 29, wherein the lipid bilayer is formed on a substantially planar electrode surface.

42. The system of claim 29, wherein the system is automated.