Sensitivity and selectivity of ion channel biosensor membranes

• US 5,234,566 A
• Filed: 04/18/1991
• Issued: 08/10/1993
• Est. Priority Date: 08/18/1988
• Status: Expired due to Term

• Current Assignee: Biosensor Enterprises, LLC
• Original Assignee: Australian Membrane and Biotechnology Research Institute
• Inventors: Raguse, Burkhard, King, Lionel G., Osman, Peter D. J., Cornell, Bruce A.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US07/654,635
• Time in Patent Office: 845 Days
• Field of Search: 204/403, 204/418, 204/416, 204/426, 435/817, 436/806
• US Class Current: 204/403.06
• CPC Class Codes: C12Q 1/002 Electrode membranes
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property

Abstract

The present invention provides a biosensor comprising at least one lipid membrane, each membrane including at least one gated ion channel. The membranes comprise a closely packed array of self-assembly amphophilic molecules and the gated ion channel has a conductance which is dependent upon an electric field applied across the membrane. The biosensor of the present invention may comprise a plurality of discrete membranes each including at least one gated ion channel. The conductance of each of the membranes is measurable independently of the conductance of the other membranes.

Claims

1. A biosensor comprising a plurality of discrete, substantially identical membranes, each membrane including at least one gated ion channel, the conductance of each membrane being measurable independently of the conductance of the other membranes, each of said membranes comprising a closely packed array of self-assembling amphophilic molecules, at least one dedicated electrode provided on one side of the membrane which cooperates with an electrode on the other side of the membrane to enable the application of an electrical potential across the membrane, the at least one gated ion channel having a conductance which is dependent upon the electric potential applied across the membrane.

2. A biosensor as claimed in claim 1 in which the ion channel is modified by incorporation or removal of polar, dipolar or polarisable groups.

3. A biosensor as claimed in claim 1 in which at least one dedicated electrode is provided on one side of each membrane which cooperates with an electrode on the other side of the each membrane to enable the application of an electric potential across the membrane, the plurality of membranes being multiplexed by multiplexing the signal applied to or measured from the respective discrete electrodes.

5. A biosensor as claimed in claim 1 in which the ion channel is selected from the group consisting of peptides capable of forming helices and aggregates thereof, podands, coronands and cyptands.

6. A biosensor as claimed in claim 5 in which the ion channel is a peptide capable of forming a helix or aggregates thereof.

7. A biosensor as claimed in claim 6 in which the ion channel is a peptide which forms a β

helix.

8. A biosensor as claimed in claim 7 in which the ion channel is gramicidin or analogs thereof.

9. A biosensor as claimed in claim 8 in which the ion channel is gramicidin A or analogs thereof.

10. A biosensor as claimed in claim 1 in which the gates ion channel can diffuse laterally within the lipid membrane.

11. A biosensor as claimed in claim 1 in which the conductance of each lipid membrane is measured by means of a high impedance address lines, a separate address line being provided to each lipid membrane and/or multiplexing the membrane.

12. A biosensor as claimed in claim 11 in which the conductance of each lipid membrane is measured by multiplexing the membranes.

13. A biosensor as claimed in claim 12 in which the membranes are serially multiplexed.

14. A biosensor as claimed in claim 12 in which the conductance measurements are made using multiplex lines of low impedance and at least one current sensing line.

15. A biosensor as claimed in claim 14 in which there is one current sensing line.

16. A biosensor as claimed in claim 1 in which the conductance of each membrane is measured by means of switching between low impedance address lines each of which supplies a signal which is measured either by a single current sensor common to all the address lines or by a number of current sensors which are electrically isolated from each other and which measure groups of address lines.

17. A biosensor as claimed in claim 1 in which the conductance of each membrane is measured by means of switching between high impedance address lines each of which supply a signal which is measured either by a single current sensor common to all the address lines or by a number of current sensors which are electrically isolated from each other and which measure groups of address lines.

18. A biosensor as claimed in claim 11 which the gated ion channels are field effect ion channels.

19. A biosensor as claimed in claim 18 in which the plurality of discrete membranes are arranged in a two dimensional array.

20. A biosensor as claimed in claim 19 in which the multiplex lines are driven from a complex signal such that in the two dimensional array each address line in one dimension has signal components which are cross modulated with signals from address lines in the other dimension by the field effect ion channel

21. A biosensor as claimed in claim 1 in which the conductance of each lipid membrane is measured by means of a high impedance address line either by using a separate amplifier for each membrane or by switching one amplifier between each membrane or by switching a number of amplifiers between a number of membranes such that each membrane is measured.

33. A biosensor as claimed in claim 21 in which the conductance of each membrane is measured by means of switching between low impedance address lines each of which supplies a signal which is measured either by a single current sensor common to all the address lines or by a number of current sensors which are electrically isolated from each other and which measure groups of address lines.

4. A biosensor comprising a plurality of discrete substantially identical membranes, each membrane including at least one gated ion channel, each of said membranes comprising a closely packed array of self-assembling amphophilic molecules, the conductance of each of said membranes being measurable independently of the conductance of the other membranes, at least one dedicated electrode provided on one side of the membrane which cooperates with an electrode on the other side of the membrane to enable the application of an electric potential across the membrane, the signal applied to or measured from the discrete membranes.

22. A biosensor as claimed in claim 4 in which the ion channel is selected from the group consisting of peptides capable of forming helices and aggregates thereof, podands, coronands and cryptands.

23. A biosensor as claimed in claim 22 in which the ion channel is a peptide capable of forming a helix or aggregates thereof.

24. A biosensor as claimed in claim 23 in which the ion channel is a peptide which forms a β

helix.

25. A biosensor as claimed in claim 24 in which the ion channel is gramicidin or analogs thereof.

26. A biosensor as claimed in claim 25 in which the ion channel is gramicidin A or analogs thereof.

27. A biosensor as claimed in claim 4 in which the grated ion channel can diffuse laterally within the lipid membrane.

28. A biosensor as claimed in claim 4 in which the conductance of each lipid membrane is measured by means of a high impedance address lines, a separate address line being provided to each lipid membrane and/or multiplexing the membranes.

29. A biosensor as claimed in claim 28 in which the conductance of each lipid membrane is measured by multiplexing the membranes.

30. A biosensor as claimed in claim 29 in which the membranes are serially multiplexed.

31. A biosensor as claimed in claim 29 in which the conductance measurements are made using multiplex lines of low impedance and at least one current sensing line.

32. A biosensor as claimed in claim 31 in which there is one current sensing line.

34. A biosensor as claimed in claim 4 in which the conductance of each membrane is measured by means of switching between high impedance address lines each of which supply a signal which is measured either by a single current sensor common to all the address lines or by a number of current sensors which are electrically isolated from each other and which measure groups of address lines.

35. A biosensor as claimed in claim 4 in which the gated ion channels are field effect ion channels.

36. A biosensor as claimed in claim 35 in which the plurality of discrete membranes are arranged in a two dimensional array.

37. A biosensor as claimed in claim 36 in which the multiplex lines are driven from a complex signal such that in the two dimensional array each address line in one dimension has signal components which are cross modulated with signals from address lines in the other dimension by the field effect ion channel.

38. A biosensor as claimed in claim 4 in which the conductance of each lipid membrane is measured by means of a high impedance address line either by using a separate amplifier for each membrane or by switching one amplifier between each membrane or by switching a number of amplifiers between a number of membranes such that each membrane is measured.