Multi-array multi-specific electrochemiluminescence testing

• US 6,673,533 B1
• Filed: 09/17/1997
• Issued: 01/06/2004
• Est. Priority Date: 03/10/1995
• Status: Expired due to Term

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Helms, Larry R., Wohlstadter, Jacob N., Wilbur, James, Martin, Mark, Fischer, Alan, Sigal, George, Guo, Liang-Hong, Billadeau, Mark A., Leland, Jon, Darvari, Ramin
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/932,110
• Time in Patent Office: 2,302 Days
• Field of Search: 204/400, 204/403, 435/4, 435/6, 435/7.1, 435/7.2, 435/287.1, 435/287.2, 436/172, 436/518, 436/806, 436/524, 436/525, 422/52, 422/58, 422/61, 422/98, 422/102, 422/82.01
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/66 electrically excited, e.g. electroluminescence
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

Materials and methods are provided for producing patterned multi-array, multi-specific surfaces for use in diagnostics. The invention provides for electrochemiluminescence methods for detecting or measuring an analyte of interest. It also provides for novel electrodes for ECL assays. Materials and methods are provided for the chemical and/or physical control of conducting domains and reagent deposition for use multiply specific testing procedures.

Claims

1. An assay electrode comprising a composite containing a polymer matrix and a multiplicity of carbon particles dispersed therein, said assay electrode having a binding domain containing a binding reagent, wherein said binding reagent is immobilized on a surface of said electrode.

2. An assay electrode as recited in claim 1, wherein the carbon particles comprise from 0.5% to 50% by weight of the composite.

3. An assay electrode as recited in claim 1, wherein the carbon particles comprise from 1% to 30% by weight of the composite.

4. An assay electrode as recited in claim 1, wherein the carbon particles comprise from 2% to 20% by weight of the composite.

5. An assay electrode as recited in claim 1, wherein said composite is molded.

6. An assay electrode as recited in claim 1, wherein said composite is extruded.

7. An assay electrode as recited in claim 1, wherein said binding reagent is covalently bound to said electrode.

8. An assay electrode as recited in claim 1, wherein said binding reagent is non-covalently bound to said electrode.

9. An assay electrode as recited in claim 1, wherein said binding reagent is directly immobilized on said electrode.

10. An assay electrode as recited in claim 1, wherein said binding reagent is indirectly immobilized on said electrode via a binding pair.

11. An assay electrode as recited in claim 1, wherein said assay electrode includes a multiplicity of binding domains.

12. An assay electrode as recited in claim 1, wherein said binding reagent is an antibody or fragment thereof, a nucleic acid, a receptor or an enzyme.

13. An assay electrode as recited in claim 1, wherein said carbon particles are fibrils.

14. An assay electrode as recited in claim 1, wherein said carbon particles are etched or chemically modified.

15. A method for the manufacture of the assay electrode of claim 1 comprising the steps of:

(a) treating said composite containing said polymer matrix and said multiplicity of carbon particles dispersed therein with a plasma, and (b) forming said binding domain on a surface of the so-treated composite, said binding domain containing said binding reagent.

16. A method as recited in claim 15, wherein two or more sequential plasma treatments are conducted.

17. A method as recited in claim 15, wherein the plasma contains an atom or compound selected from the group consisting of O₂, Ar, H₂O, N₂, NH₃, CF₄, SF₆, C₂F₆, CHF₃, CF₂Cl₂, CF₃Br, CF₃Cl and combinations thereof.

18. A method as recited in claim 15, wherein said plasma treatment is for one or more of the purposes of:

(a) etching said polymer, or (b) derivatizing the exposed surfaces of the carbon particles dispersed in said polymer.

19. A method as recited in claim 15, wherein said carbon particles are fibrils.

20. A method as recited in claim 15, wherein said binding domain is formed by introducing said binding reagent through a hole in a mask placed on said composite.

21. An assay electrode prepared by the method claim 15.

22. The method of claim 15, wherein said multiplicity of carbon particles includes exposed carbon particles.

23. A method for the manufacture of the assay electrode of claim 1 comprising the steps of:

(a) treating said composite containing said polymer matrix and said multiplicity of carbon particles dispersed therein with a chemical reagent, and

(b) forming said binding domain on a surface of the so-treated composite, said binding domain containing said binding reagent.

24. A method as recited in claim 23, wherein said carbon particles are fibrils.

25. A method as recited in claim 23, wherein said chemical reagent is an oxidizing agent.

26. A method as recited in claim 23, wherein said binding domain is formed by introducing said binding reagent through a hole in a masked placed on said composite.

27. An assay electrode prepared by the method of claim 23.

28. The assay electrode of claim 1, wherein said multiplicity of carbon particles includes exposed carbon particles.

29. The assay electrode of claim 1, wherein said composite is plasma etched.

30. A cartridge for use in an instrument system for conducting electrochemiluminescence assays for the detection or quantitation of an analyte, comprising:

one or more electrodes of claim 1 and assay reagents.

31. A cartridge as recited in claim 30, which does not contain a liquid assay reagent.

32. A cartridge as recited in claim 30, wherein said assay reagents include a dry assay reagent.

33. A cartridge as recited in claim 32, wherein said dry assay reagent is an electrochemiluminescence coreactant.

34. A cartridge as recited in claim 32, wherein said dry assay reagent is a buffer.

35. A cartridge as recited in claim 32, wherein said dry assay reagent contains an electrochemiluminescent moiety.

36. A cartridge as recited in claim 32, wherein said dry assay reagent is a calibration standard.

37. A cartridge as recited in claim 32, wherein said dry assay reagent is a preservative.

38. A cartridge as recited in claim 32, wherein said dry assay reagent is a carbohydrate.

39. A cartridge as recited in claim 30, wherein said electrode defines an interior wall of a cell for containing a liquid sample.

40. A cartridge as recited in claim 30, including a window for passage of light from said cartridge to a light detector for detecting light from said assay.

41. A cartridge as recited in claim 30, including fluid metering means.

42. A cartridge as recited in claim 30, including means for modulating the temperature of said cartridge.

43. A cartridge as recited in claim 30, including means for determining the temperature of said cartridge.

44. A cartridge as recited in claim 30, including a fluid path.

45. A cartridge as recited in claim 30, including a counter electrode.

46. A cartridge as recited in claim 30, including a reference electrode.

47. A cartridge as recited in claim 30, including a source of electrical energy.

48. A cartridge as recited in claim 30, including means for containment of a fluid sample introduced to said cartridge.

49. A cartridge as recited in claim 30, which is injection molded.

50. A cartridge as recited in claim 30, which is disposable.

51. A cartridge as recited in claim 30, including means for mixing contents thereof.

52. A cartridge as recited in claim 51, wherein said means for mixing is a sonication device.

53. A cartridge as recited in claim 52, wherein said sonication device is a piezoelectric device structurally coupled to the electrode in said cartridge.

54. An instrument system for conducting electrochemiluminescence assays for the detection or quantitation of an analyte comprising:

(a) a cartridge including one or more assay electrodes of claim 1; and

(b) a light detector for selectively detecting or quantitating light from reach of said binding domains.

55. An instrument system as recited in claim 54, wherein said carbon particles are fibrils.

56. An instrument system as recited in claim 54, including means for mixing contents of said cartridge.

57. An instrument system as recited in claim 54, wherein said cartridge includes a dry assay reagent.

58. An instrument as recited in claim 54, wherein said light detector is a CCD.

59. An instrument as recited in claim 54, wherein said light detector is a photodiode.

60. An instrument as recited in claim 54, including temperature control means for controlling the temperature of said cartridge.

61. An instrument system for conducting electrochemilumnescence assays for the detection or quantitation of an analyte comprising:

(a) a cartridge including one or more electrodes of claim 1;

(b) means for selectively detecting or quantitating light from each of said binding domains; and

(c) means for mixing contents of said cartridge.

62. An instrument system as recited in claim 61, wherein said mixing system is a sonication device.

63. An apparatus for use in the detection of an analyte comprising the assay electrode of claim 1 and a source of electrical energy, wherein said multiplicity of carbon particles include carbon fibrils.

64. A cartridge as recited in claim 63, containing in addition to said one or more electrodes, a dry assay reagent.

65. An assay electrode comprising a composite containing a matrix and a multiplicity of carbon fibrils dispersed therein, said electrode having a binding domain containing an assay reagent on a surface thereof.

66. A cartridge for use in an instrument system for conducting electrochemiluminescence assays for the detection or quantitation of an analyte, comprising:

one or more assay electrodes of claim 65.

67. A solid phase support comprising:

a composite containing a matrix and a multiplicity of carbon fibrils dispersed therein, wherein said support further comprises one or more binding domains on a surface thereof containing binding reagents.

68. A solid phase support as recited in claim 67, wherein said matrix is a polymer.

69. A solid phase support as recited in claim 67, wherein said support is used in the detection or quantitation of analyte.

70. A solid phase support as recited in claim 67, further comprising an enzyme immobilized on said support.

71. The solid phase support of claim 67, further comprising a biomolecule immobilized in said binding domain.

72. The solid phase support of claim 71, wherein said biomolecule is attached to said binding domain covalently.

73. The solid phase support of claim 71, wherein said biomolecule is attached to said binding domain non-covalently.

74. The solid phase support of claim 71, wherein said biomolecule is labeled with an electrochemiluminescent moiety.

75. The solid phase support of claim 71, wherein said biomolecule is bound to a second biomolecule labeled with an electrochemiluminescent moiety via one or more specific binding interactions.

76. The solid phase support of claim 71, where said biomolecule comprises an antibody, nucleic acid, an enzyme or a peptide.

77. The solid phase support of claim 71, wherein said support is coupled to a sonication device.

78. The solid phase support of claim 71, wherein said composite comprises 1% to 30% carbon fibrils.

79. The solid phase support of claim 71, wherein at least a portion said carbon fibrils are exposed by chemical oxidation.

80. The solid phase support of claim 71, wherein at least a portion said carbon fibrils are exposed by plasma oxidation.

81. An assay electrode comprising a matrix and a multiplicity of carbon fibrils dispersed therein, said assay electrode having a biomolecule immobilized on a surface thereof.

82. An assay electrode comprising a composite containing a polymer matrix and a multiplicity of carbon fibrils dispersed therein, said assay electrode having a binding domain containing a reagent on a surface thereof.

83. The electrode of claim 82, wherein said reagent is attached to said binding domain covalently.

84. The electrode of claim 82, wherein said reagent is attached to said binding domain non-covalently.

85. The electrode of claim 82, wherein said reagent is labeled with an electrochemiluminescent moiety.

86. The electrode of claim 82, wherein said reagent is bound to a second reagent labeled with an eletrochemiluminescent moiety via one or more specific binding interactions.

87. The electrode of claim 82, wherein said reagent comprises an antibody, nucleic acid, an enzyme or a peptide.

88. The electrode of claim 82, wherein said electrode is coupled to a sonication device.

89. The electrode of claim 82, wherein said composite comprises 1% to 30% carbon fibrils.

90. The electrode of claim 82, wherein at least a portion of said carbon fibrils is exposed by chemical oxidation.

91. The electrode of claim 82, wherein at least a portion of said carbon fibrils is exposed by plasma oxidation.

92. An assay electrode comprising a composite containing a polymer matrix having exposed carbon particles, said assay electrode having a binding domain containing a binding reagent on a surface thereof.