Assay electrode having immobilized lipid/protein layers, methods of making the same and methods of using the same for luminescence test measurements

• US 20030124572A1
• Filed: 07/30/2002
• Published: 07/03/2003
• Est. Priority Date: 07/30/2001
• Status: Active Grant

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Favor of MESO Scale Technologi
• Inventors: Mathew, Anu, Maheshwari, Gargi, Umek, Robert M., Eason, Paula Denney, Woods, Douglas
• Granted Patent: US 8,815,577 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/66 electrically excited, e.g. electroluminescence
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

Improved systems, system components, and methods for performing assays involving biological membranes and/or components thereof. Preferred luminescence test measurements are conducted using an assay module with integrated electrodes having biological membranes and/or components thereof immobilized thereon with a reader apparatus adapted to receive assay modules, induce luminescence, preferably electrode induced luminescence, in the wells or assay regions of the assay modules and measure the induced luminescence.

Claims

1. An assay module comprising at least one electrode having at least one biomaterial passively adsorbed thereon, wherein said biomaterial is selected from the group consisting of membrane protein, lipid/protein layer, and/or membrane lipid.

36. A kit for use in conducting electrode induced luminescence assays comprising, in one or more containers, the assay module of claim 1 and at least one assay component selected from the group consisting of:

(a) at least one luminescent label;

(b) at least one luminescence coreactant;

(c) one or more binding reagents;

(d) a pH buffer;

(e) one or more blocking reagents;

(f) preservatives;

(g) stabilizing agents;

(h) enzymes;

(i) detergents; and

(j) desiccants.

2. An assay module comprising at least one electrode, said electrode comprising elemental carbon and having at least one biomaterial immobilized thereon, wherein said biomaterial is selected from the group consisting of membrane protein, lipid/protein layer, and/or membrane lipid.

3. The assay module of claim 2, wherein said biomaterial is selected from the group consisting of intact cell, cell lysate, cell membrane fragment, cell membrane, membrane ghost, organelle, organelle membrane fragment, organelle membrane, virion, virion fragment, virion membrane, liposome, detergent solubilized protein, detergent solubilized lipid or combinations thereof.

4. The assay module of claim 2, wherein said biomaterial is selected from the group consisting of intact cell, cell membrane and cell membrane fragment.

5. The assay module of claim 2, wherein said electrode comprises carbon ink.

6. The assay module of claim 2, wherein said electrode is a screen-printed carbon ink.

7. The assay module of claim 2, wherein said biomaterial comprises a lipid/protein layer which contains integral membrane proteins.

8. The assay module of claim 2, wherein said biomaterial comprises a lipid/protein layer which contains peripheral proteins.

9. The assay module of claim 2, wherein said biomaterial comprises between 0.3 micrograms to 30 micrograms of total protein per 1 in electrode.

10. The assay module of claim 2, wherein said biomaterial is passively adsorbed on said electrode.

11. The assay module of claim 2, wherein said biomaterial is covalently attached to said electrode.

12. The assay module of claim 2, wherein said biomaterial is in a dry state.

13. The assay module of claim 2, wherein said biomaterial is fixed on said electrode.

14. The assay module of claim 13, wherein said biomaterial is immobilized via a specific binding reaction with a binding partner that is affixed on said electrode.

15. The assay module of claim 14, wherein said binding partner is passively adsorbed on said electrode.

16. The assay module of claim 14, wherein said binding partner is covalently attached to said electrode.

17. The assay module of claim 14, wherein said binding partner is affixed to said electrode via a second bio-specific interaction with a second binding partner.

18. The assay module of claim 17, wherein said second bio-specific interaction is selected from the group consisting of biotin-avidin, biotin-streptavidin, and antibody-hapten.

19. The assay module of claim 13, wherein said biomaterial is immobilized onto a streptavidin surface or an avidin surface on said electrode via a biotin:

streptavidin or biotin;

avidin linkage.

20. The assay module of claim 13, wherein said biomaterial is immobilized onto an antibody surface on said electrode via a specific binding reaction of antibodies in said antibody surface with a constituent of said biomaterial.

21. The assay module of claim 13, wherein said biomaterial is immobilized onto a lectin surface via a specific binding interaction of lectins in said lectin surface with a constituent of said biomaterial.

22. The assay module of claim 21, wherein said lectin surface comprises a lectin selected from the group consisting of phytohemagglutinin, pokeweed mitogen, wheat gern agglutinin and concanavalin A.

23. The assay module of claim 22, wherein said lectin surface comprises phytohemagglutinin.

24. The assay module of claim 22, wherein said lectin surface comprises pokeweed mitogen.

25. The assay module of claim 13, wherein a surface of said electrode is coated with microtubules and said biomaterial is immobilized onto said coated surface.

26. The assay electrode of claim 2, wherein said assay electrode comprises an array of biomaterials immobilized thereon.

27. An assay module comprising a plurality of patterned electrodes, said patterned electrodes having at least one biomaterial immobilized thereon, wherein said biomaterial is selected from the group consisting of membrane protein, lipid/protein layer, and/or membrane lipid.

28. The assay module of claim 27, wherein said biomaterial is selected from the group consisting of intact cell, cell lysate, cell fragment, cell membrane, membrane ghost, organelle, organelle fragment, organelle membrane, virion, virion fragment, virion membrane, liposome, detergent solubilized protein, detergent solubilized lipid or combinations thereof.

29. The assay module of claim 27, wherein said biomaterial is selected from the group consisting of intact cell, cell membrane and cell membrane fragment.

30. The assay module of claim 27, wherein at least two of said patterned electrodes have different biomaterials immobilized thereon.

31. A multi-well plate having a plurality of wells, said wells comprising at least one electrode within said wells and said at least one electrode having at least one membrane protein, at least one lipid/protein layer and/or at least one membrane lipid immobilized thereon.

32. The multi-well plate of claim 31, wherein said electrode comprises elemental carbon.

33. The multi-well plate of claim 31, wherein said at least one electrode comprises a working electrode surface bounded by a dielectric surface and having at least one membrane protein, at least one lipid/protein layer, and/or at least one membrane lipid immobilized thereon.

34. The multi-well plate of claim 31, wherein said at least one electrode comprises a working electrode having one or more assay domains comprising one or more immobilized lipid proteins, one or more lipid/protein layers and/or one or more membrane lipids, said assay domains being defined by openings in one or more dielectric layers supported on said working electrode.

35. The multi-well plate of claim 31, wherein said at least one electrode comprises a plurality of working electrodes having assay domains comprising one or more immobilized lipid proteins, one or more lipid/protein layers and/or one or more membrane lipids, said assay domains being defined by openings in one or more dielectric layers supported on said working electrode.

37. A method of immobilizing a biomaterial on an electrode comprising elemental carbon, said method comprising the steps of:

a. affixing said biomaterial on said electrode and

b. drying said electrode

wherein said biomaterial is selected from the group consisting of membrane protein, lipid/protein layer, and/or membrane lipid.

38. The method of claim 37, wherein said affixing comprises adsorbing said biomaterial by passive adsorption.

39. The method of claim 37, wherein said affixing comprises binding said biomaterial via a biospecific interaction with a binding partner immobilized on said electrode.

40. A method of measuring a binding interaction of a biomaterial selected from membrane proteins, lipid/protein layers and/or membrane lipids, said method comprising:

(a) contacting one or more electrodes having one or more of said biomaterials immobilized thereon with one or more binding partners of said one or more biomaterials; and

(b) measuring binding between said one or more biomaterials and said one or more binding partners using electrode induced luminescence.

41. The method of claim 40, wherein said one or more binding partners are labeled with electrochemiluminescent labels.

42. The method of claim 40, wherein said electrode induced luminescence is induced by a applying electrical energy to said one or more electrodes.

43. The method of claim 40, wherein said method further comprises the step of contacting the biomaterial and/or the binding partner with an inhibitor of the binding interaction and thereby measuring inhibition of said binding interaction.

58. The method of claim 40, wherein said electrode induced luminescence is induced in the presence of a solution comprising an electrochemiluminescence coreactant.

59. The method of claim 58, wherein said coreactant is a trialkylamine.

60. The method of claim 58, wherein said solution does not comprise a surfactant.

61. The method of claim 58, wherein said solution comprises a surfactant.

44. A method of measuring an activity or a process that modifies a biomaterial and/or a binding partner of the biomaterial, said biomaterial selected from membrane proteins, lipid/protein layers and/or membrane lipids, said method comprising:

(a) subjecting said biomaterial and/or said binding partner to said activity and/or said process conditions; and

(b) measuring binding between said binding partner and said biomaterial using electrode induced luminescence while said biomaterial is immobilized on one or more electrodes.

45. The method of claim 44, wherein said biomaterial and/or said binding partner is subjected to said activity and/or said process prior to immobilizing said biomaterial.

46. The method of claim 44, wherein said biomaterial and/or said binding partner is subjected to said activity and/or said process after immobilizing said biomaterial.

47. The method of claim 44, wherein said biomaterial and/or said binding partner is subjected to said activity and/or said process in situ during immobilizing of said biomaterial.

48. The method of claim 44, wherein step (a) comprises subjecting said biomaterial and/or said binding partner to an inhibitor of said process or activity.

62. The method of claim 44, wherein said electrode induced luminescence is induced in the presence of a solution comprising an electrochemiluminescence coreactant.

63. The method of claim 62, wherein said coreactant is a trialkylamine.

64. The method of claim 62, wherein said solution does not comprise a surfactant.

65. The method of claim 62, wherein said solution comprises a surfactant.

49. A method for measuring a binding interaction or biomaterial activity comprising:

(a) contacting a plurality of discrete lipid proteins, lipid/protein layers and/or membrane lipids immobilized on an electrode with one or more binding partners of said lipid proteins, lipid/protein layers and/or membrane lipids;

(b) applying a voltage waveform effective to trigger luminescence at said electrode in the presence of a reaction medium suitable for conducting said electrode induced luminescence assay; and

(c) measuring emitted luminescence.

66. The method of claim 49, wherein said electrode induced luminescence is induced in the presence of a solution comprising an electrochemiluminescence coreactant.

67. The method of claim 66, wherein said coreactant is a trialkylamine.

68. The method of claim 66, wherein said solution does not comprise a surfactant.

69. The method of claim 66, wherein said solution comprises a surfactant.

50. An assay module comprising a surface having pokeweed mitogen immobilized thereon.

51. A multi-well plate, wherein at least one well of said plate has pokeweed mitogen immobilized on a surface thereon.

52. A method of immobilizing a biomaterial comprising contacting the biomaterial with a surface having pokeweed mitogen affixed thereon.

53. A method of conducting a magnetic particle based electrochemiluminescence assay wherein said magnetic particle comprises an immobilized layer of a biomaterial selected from the group consisting of membrane protein, lipid/protein layer, and/or membrane lipid.

54. The method of claim 53, wherein said electrode induced luminescence is induced in the presence of a solution comprising an electrochemiluminescence coreactant.

55. The method of claim 54, wherein said coreactant is a trialkylamine.

56. The method of claim 54, wherein said solution does not comprise a surfactant.

57. The method of claim 54, wherein said solution comprises a surfactant.