IP Library Granted Patent US 12,042,790
Granted Patent B2
US 12,042,790 · App. 16/463,617 · Granted Jul 23, 2024

Apparatus and methods for controlling insertion of a membrane channel into a membrane

Inventors: Daniel Ryan Garalde (Cambridge, MA); James Anthony Clarke (Oxford, GB); Mike Jennison (Oxford, GB); Andrew John Heron (Oxford, GB)
Assignee: Oxford Nanopore Technologies PLC
B01L3/502B81C1/00071B01L2300/044B01L2400/0415
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Quick Facts
Patent No.
US 12,042,790
App. No.
16/463,617
Granted
Jul 23, 2024
Kind
B2
Abstract

Apparatus and methods for controlling the insertion of a membrane channel into a membrane are disclosed. In one arrangement a first bath holds a first liquid in contact with a first surface of a membrane. A second bath holds a second liquid in contact with a second surface of the membrane. The membrane separates the first and second liquids. A first electrode contacts the first liquid. A second electrode contacts the second liquid. A driving unit applies a potential difference across the membrane via the first and second electrodes to promote insertion of a membrane channel into the membrane from the first liquid or the second liquid. A membrane voltage reduction unit is connected in series with the membrane. The driving unit applies a driving voltage across the membrane voltage reduction unit and the membrane, the driving voltage providing the potential difference across the membrane. The membrane voltage reduction unit is configured such that a reduction in resistance through the membrane caused by insertion of a membrane channel intrinsically increases a potential difference across the membrane voltage reduction unit thereby lowering the potential difference across the membrane. The lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel.

Claims (45)

1. An apparatus for controlling insertion of a membrane channel into a membrane, comprising:

a first bath for holding a first liquid in contact with a first surface of the membrane;

a second bath for holding a second liquid in contact with a second surface of the membrane, wherein the membrane separates the first and second liquids;

a first electrode configured to contact the first liquid;

a second electrode configured to contact the second liquid; and

a driving unit configured to apply a potential difference across the membrane via the first and second electrodes to promote insertion of a membrane channel into the membrane from the first liquid or the second liquid, wherein:

the apparatus comprises a membrane voltage reduction unit connected in series with the membrane;

the driving unit is configured to apply a driving voltage across the membrane voltage reduction unit and the membrane, the driving voltage providing the potential difference across the membrane; and

the membrane voltage reduction unit is configured such that a reduction in resistance through the membrane caused by insertion of a membrane channel increases a potential difference across the membrane voltage reduction unit thereby lowering the potential difference across the membrane, wherein the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel.

2. The apparatus of claim 1 , wherein the membrane voltage reduction unit is configured such that the lowering of the potential difference across the membrane is triggered without logic control.

3. The apparatus of claim 1 , wherein:

the membrane voltage reduction unit comprises a current source configured to supply a constant current through resistances up to a maximum resistance; and

the lowering of the potential difference across the membrane is caused by the resistance through the membrane being reduced from a resistance above said maximum resistance to a resistance below said maximum resistance.

4. The apparatus of claim 1 , wherein:

the membrane voltage reduction unit comprises a resistive component in series with the membrane, wherein the resistance of the resistive component is selected to ensure that the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel, while also allowing the potential difference across the membrane to be sufficiently high prior to insertion to promote insertion of the membrane channel prior to insertion.

5. The apparatus of claim 1 , wherein:

the membrane voltage reduction unit comprises a diode in series with the membrane, wherein the diode is configured such that the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel, while also allowing the potential difference across the membrane to be sufficiently high prior to insertion to promote insertion of the membrane channel prior to insertion.

6. The apparatus of claim 1 , wherein the driving voltage is increased from a minimum driving voltage to a predetermined maximum driving voltage.

7. The apparatus of claim 6 , wherein the increase in driving voltage from a minimum to a maximum value is completed in less than 1 s.

8. The apparatus of claim 1 , wherein:

a plurality of the second baths are provided, each second bath being configured to support a different membrane;

a plurality of the membrane voltage reduction units are provided, each membrane voltage reduction unit being connected in series with a different membrane or a different portion of the same membrane; and

the driving unit is configured to apply the driving voltage in parallel across all of the pairs of membrane voltage reduction unit and different membrane or across all of the pairs of membrane voltage reduction unit and different portion of the same membrane.

9. The apparatus of claim 1 , wherein the membrane comprises an amphiphilic membrane.

10. A method of controlling insertion of a membrane channel into a membrane, comprising:

providing a membrane in contact with and separating first and second liquids on respective first and second sides of the membrane;

using a driving unit to apply a potential difference across the membrane via first and second electrodes to promote insertion of a membrane channel into the membrane from the first liquid or the second liquid, wherein:

the driving unit comprises a membrane voltage reduction unit connected in series with the membrane;

the driving unit is configured to apply a driving voltage across the membrane voltage reduction unit and the membrane, the driving voltage providing the potential difference across the membrane; and

the membrane voltage reduction unit is configured such that a reduction in resistance through the membrane caused by insertion of the membrane channel increases a potential difference across the membrane voltage reduction unit thereby lowering the potential difference across the membrane, wherein the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel.

11. The method of claim 10 , wherein the membrane voltage reduction unit is configured such that the lowering of the potential difference across the membrane is triggered without computer control.

12. The method of claim 10 , wherein:

the membrane voltage reduction unit comprises a current source capable of supplying a predetermined constant current through resistances up to a maximum resistance; and

the lowering of the potential difference across the membrane is caused by the resistance through the membrane being reduced from a resistance above said maximum resistance to a resistance below said maximum resistance.

13. The method of claim 10 , wherein:

the membrane voltage reduction unit comprises a resistive component in series with the membrane, wherein the resistance of the resistive component is selected to ensure that the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel, while also allowing the potential difference across the membrane to be sufficiently high prior to insertion to promote insertion of the membrane channel prior to insertion.

14. The method of claim 10 , wherein:

the membrane voltage reduction unit comprises a diode in series with the membrane, wherein the diode is configured such that the lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel, while also allowing the potential difference across the membrane to be sufficiently high prior to insertion to promote insertion of the membrane channel prior to insertion.

15. The method of claim 10 , wherein the driving voltage is progressively increased from zero voltage or a minimum driving voltage to a predetermined maximum driving voltage.

16. The method of claim 15 , wherein the progressive increase is completed in less than 5 s.

17. The method of claim 10 , wherein:

a plurality of the second baths are provided, each second bath being configured to support a different membrane or a different portion of the same membrane;

a plurality of the membrane voltage reduction units are provided, each membrane voltage reduction unit being connected in series with a different membrane or a different portion of the same membrane; and

the driving unit is configured to apply the driving voltage in parallel across all of the pairs of membrane voltage reduction unit and different membrane or across all of the pairs of membrane voltage reduction unit and different portion of the same membrane.

18. The method of claim 10 , wherein the membrane comprises an amphiphilic membrane.

Assignments (2)
CHANGE OF NAME Recorded Jan 14, 2022
From: OXFORD NANOPORE TECHNOLOGIES LIMITED
To: OXFORD NANOPORE TECHNOLOGIES PLC
Reel/Frame 058737/0664 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 31, 2020
From: GARALDE, DANIEL RYAN; CLARKE, JAMES ANTHONY; JENNISON, MIKE; HERON, ANDREW JOHN
To: OXFORD NANOPORE TECHNOLOGIES LIMITED
Reel/Frame 052274/0001 →
Priority Claims (1)
GB 1619930 · Nov 24, 2016 · national
Continuity (1)
Related Publication 20200179920A1 · Jun 11, 2020
Cited By (1)
US 12,515,215