Receptor membranes

• US 5,766,960 A
• Filed: 05/23/1995
• Issued: 06/16/1998
• Est. Priority Date: 07/27/1987
• Status: Expired due to Term

• Current Assignee: Biosensor Enterprises, LLC
• Original Assignee: Australian Membrane and Biotechnology Research Institute
• Inventors: Braach-Maksvytis, Vijoleta Lucija Bronislava, Cornell, Bruce Andrew
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Swartz, Rodney P.
• Application Number:
  US08/449,895
• Time in Patent Office: 1,120 Days
• Field of Search: 435/71, 435/317.1, 435/817, 436/501, 436/512, 436/518, 436/524, 436/525, 436/532, 436/547
• US Class Current: 436/501
• CPC Class Codes: B01D 69/02 characterised by their properties
• CPC Class Codes: B01D 69/122 Separate manufacturing of ultra-thin membranes
• CPC Class Codes: B01D 69/141 Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 33/5432 Liposomes or microcapsules
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

A membrane comprising a closely packed array of self-assembling amphiphilic molecules, and is characterized in that it incorporates a plurality of ion channels, and/or at least a proportion of the self-assembling molecules comprise a receptor molecule conjugated with a supporting entity. The ion channel is selected from the group consisting of peptides capable of forming helices and aggregates thereof, corronands, cryptands, podands and combinations thereof. In the amphiphilic molecules comprising a receptor molecule conjugated with a supporting entity, the receptor molecule has a receptor site and is selected from the group consisting of immunoglobulins, antibodies, antibody fragments, dyes enzymes and lectins. "The supporting entity is selected from the group consisting of a lipid head group, a hydrocarbon chain(s), a cross-linkable molecule and a membrane protein. The supporting entity is attached to the receptor molecule at tan end remote from the receptor site. In preferred embodiments the ion channel is gramicidin A, and is preferable gated. Such membranes may be used in the formation of sensing devices.

Claims

1. A membrane comprising a closely packed array of self-assembling amphiphilic molecules, the membrane being characterized in the (1) the membrane includes a plurality of ion channels selected from the group consisting of podands, cryptands and coronands:

(2) at least a proportion of the self assembling amphiphilic molecules comprise a receptor molecule conjugated with a supporting entity, the supporting entity being conjugated with the receptor molecule at a location from the receptor site.

2. A membrane as claimed in claim 1 in which a receptor moiety is attached or bound to the ion channel at an end thereof, the receptor moiety being such that it normally exists in a first state, but when bound to an analyte exists in a second state, said change of state of the receptor moiety causing a change in the ability of ions to pass through the ion channel.

3. A membrane as claimed in claim 2 in which the receptor moiety is an antibody or antibody binding fragments.

4. A membrane as claimed in claim 1 in which the receptor molecule is an antibody or antibody binding fragments.

5. A membrane as claimed in claim 4 in which the antibody binding fragment is attached to a hydrocarbon chain(s) and in which up to 2% of the amphiphilic molecules in the membrane comprise an Fab fragment attached to a hydrocarbon chain(s).

6. A membrane as claimed in claim 1 in which a proportion of amphiphilic molecules comprise a receptor molecule conjugated with a supporting entity, individual receptor molecules being reactive with different molecules or antigenic determinants.

7. A membrane as claimed in claim 1 in which an antibody or antibody fragment is attached to the ion channel such that when an antigen is bound to the antibody or antibody fragment ions can pass through the ion channel or are prevented from passing through the channel.

8. A membrane as claimed in claim 1 in which the amphiphilic molecules, not including a receptor molecule, include or are decorated with at least one moiety cross-linked with at least one corresponding moiety on another of these molecules.

9. A membrane as claimed in claim 1 in which the ion channels and/or the proportion of amphiphilic molecules comprising the receptor molecule conjugated with the supporting entity, each include or are decorated with at least one moiety cross-linked with at least one corresponding moiety on another molecule.

10. A membrane as claimed in claim 1 in which the membrane is attached to a solid surface, the attachment being by means of groups provided on the membrane, said groups reactive with the solid surface or groups provided thereon.

11. A membrane as claimed in claim 2 in which the receptor moiety is a plugging compound which plugs the ion channel or in which the plugging compound which plugs the ion channel is attached to the receptor moiety, the bind of the analyte to the receptor moiety causing a change in the relationship between the plugging compound and the ion channel and altering the ability of ions to pass through the ion channel.

12. A membrane as claimed in claim 11 in which the plugging compound is a positively charged species with an ionic diameter of 4 to 6 Angstroms.

35. A membrane as claimed in claim 2 in which the membrane is a bilayer.

38. A membrane as claimed in claim 2 in which the membrane is a monolayer.

41. A membrane as claimed in claim 2 in which a proportion of the amphiphilic molecules are membrane spanning lipids.

44. A membrane as claimed in claim 2 in which the receptor moiety is attached to the ion channel via a non-hydrophobic linker group.

47. A membrane as claimed in claim 2 in which the receptor moiety is attached to the supporting entity via a non-hydrophobic linker group.

49. A membrane as claimed in claim 2 in which the receptor moiety is attached to the supporting entity via a non-hydrophobic linker group.

51. The membrane of claim 1, wherein the receptor molecule is selected from the group consisting of antigens, immunoglobulins, peptides, antibodies, antibody binding fragments, enzymes, dyes, and lectins.

13. A membrane comprising a closely packed array of self-assembling amphiphilic molecules, the membrane including a plurality of ion channels selected from the group consisting of podands, cryptands and coronands, a receptor moiety being attached to the ion channel at an end thereof, the receptor moiety being such that it normally exists in a first state, but when bound to an analyte exists in a second state, said change of state of the receptor moiety causing a change in the ability of ions to pass through the ion channel.

14. A membrane as claimed in claim 13 in which the receptor moiety is an antibody or antibody binding fragments.

15. A membrane as claimed in claim 13 in which the amphiphilic molecules and/or the ion channels each include or are decorated with at least one moiety cross-linked with at least one corresponding moiety on another molecule.

16. A membrane as claimed in claim 13 in which the membrane is attached to a solid surface, the attachment being by means of groups provided on the membrane, said groups being reactive with the solid surface or groups provided thereon.

17. A membrane as claimed in claim 13 in which the receptor moiety is a plugging compound which plugs the ion channel or in which a plugging compound which plugs the ion channel is attached to the receptor moiety, the binding of the analyte to the receptor moiety causing a change in the relationship between the plugging compound and the ion channel and causing a change in the ability of ions to pass through the ion channel.

18. A membrane as claimed in claim 17 in which the plugging compound is apositively charged species with an ionic diameter of 4 to 6 Angstroms.

36. A membrane as claimed in claim 13 in which the membrane is a bilayer.

39. A membrane as claimed in claim 13 in which the membrane is a monolayer.

42. A membrane as claimed in claim 13 in which a proportion of the amphiphilic molecules are membrane spanning lipids.

45. A membrane as claimed in claim 13 in which the receptor moiety is attached to the ion channel via a non-hydrophobic linker group.

52. The membrane of claim 13, wherein the receptor moiety is selected from the group consisting of antigens, immunoglobulins, peptides, antibodies, antibody binding fragments, enzymes, dyes, and lectins.

19. A membrane comprising a closely packed array of self-assembling amphiphilic molecules, the membrane being characterized in that (1) the membrane includes a plurality of ion channels selected from the group consisting of peptides capable of forming helices and aggregates thereof;

and (2) at least a proportion of the self assembling amphiphilic molecules comprise a receptor molecule conjugated with a supporting entity, the receptor molecule having a receptor site, the supporting entity being selected from the group consisting of hydrocarbon chains having lipid head groups and cross-linkable molecules, the supporting entity being conjugated with the receptor molecule at a location remote from the receptor site.

20. A membrane as claimed in claim 19 in which the ion channels are aggregates of α

helical peptides.

21. A membrane as claimed in claim 19 in which the ion channels are peptides which form a β

helix.

22. A membrane as claimed in claim 21 in which the ion channels are gramicidin or analogues thereof.

23. A membrane as claimed in claim 21 in which the ion channel is gramicidin A or analogues thereof.

24. A membrane as claimed in claim 19 in which a receptor moiety is attached or bound with the ion channel at an end thereof, the receptor moiety being such that it normally exists in a first state, but when bound to an analyte exists in a second state, said change of state of the receptor moiety causing a change in the ability of ions to pass through the ion channel.

25. A membrane as claimed in claim 24 in which the receptor moiety is an antibody or antibody binding fragments.

26. A membrane as claimed in claim 19 in which the receptor molecule is an antibody or antibody binding fragments.

27. A membrane as claimed in claim 19 in which a proportion of amphiphilic molecules comprise a receptor molecule conjugated with a supporting entity, individual receptor molecules being reactive with different molecules or antigenic determinants.

28. A membrane as claimed in claim 19 in which an antibody or antibody fragment is attached to the ion channel such that when an antigen is bound to the antibody or antibody binding fragments the ability of ions to pass through the ion channel is altered.

29. A membrane as claimed in claim 19 in which the amphiphilic molecules, not including a receptor molecule, include or are decorated with at least one moiety cross-linked with at least one corresponding moiety on another of these molecules.

30. A membrane as claimed in claim 19 in which the ion channels and/or the proportion of amphiphilic molecules comprising the receptor molecule conjugated with the supporting entity, each include or are decorated with at least one moiety cross-linked with at least one corresponding moiety on another molecule.

31. A membrane as claimed in claim 19 in which the membrane is attached to a solid surface, the attachment being by means of groups provded on the membrane, said groups reactive with the solid surface or groups provided thereon.

32. A membrane as claimed in claim 24 in which the receptor moiety is a plugging compound which plugs the ion channel or in which a plugging compound which plugs the ion channel is attached to the receptor moiety, the binding of the analyte to the receptor moiety causing a change in the relationship between the plugging compound and the ion channel and causing a change in the ability of ions to pass through the ion channel.

33. A membrane as claimed in claim 32 in which the plugging compound is a positively charged species with an ionic diameter of 4 to 6 Angstroms.

34. A membrane as claimed in claim 24 in which the ion channel is dimeric gramicidin A and the receptor moiety is an antibody fragment, the ability of ions to pass through the ion channel being altered upon the binding of the antibody fragment to an analyte due to the disruption of the dimeric gramicidin A backbone, or to the disruption of the portion of the helix of the dimeric gramicidin attached to an Fab fragment.

37. A membrane as claimed in claim 19 in which the membrane is a bilayer.

40. A membrane as claimed in claim 19 in which the membrane is a monolayer.

43. A membrane as claimed in claim 19 in which a proportion of the amphiphilic molecules are membrane spanning lipids.

46. A membrane as claimed in claim 24 in which the receptor moiety is attached to the ion channel via a non-hydrophobic linker group.

48. A membrane as claimed in claim 19 in which the receptor moiety is attached to the supporting entity via a non-hydrophobic linker group.

50. A membrane as claimed in claim 24 in which the receptor moiety binds to the receptor molecule.

53. The membrane of claim 19, wherein the receptor molecule is selected from the group consisting of antigens, immunoglobulines, peptides, antibodies, antibody binding fragments, enzymes, dyes, and lectins.