Multi-array, multi-specific electrochemiluminescence testing

• US 6,066,448 A
• Filed: 03/06/1996
• Issued: 05/23/2000
• Est. Priority Date: 03/10/1995
• Status: Expired due to Term

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Wohlstadter, Jacob N., LeLand, Jon, Wilbur, James, Martin, Mark, Fischer, Alan, Sigal, George, Guo, Liang-Hong
• Primary Examiner(s): CHIN, CHRISTOPHER L
• Application Number:
  US08/611,804
• Time in Patent Office: 1,539 Days
• Field of Search: 435/4, 435/6, 435/7.1, 435/7.2, 435/287.1, 435/287.2, 436/172, 436/518, 436/806, 422/52, 422/58, 422/61, 422/98, 422/102, 422/82.01, 204/400, 204/403
• US Class Current: 435/6.11
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 977/957 Of chemical property or presence

Abstract

Materials and methods are provided for producing patterned multi-array, multi-specific surfaces which are electronically excited for use in electrochemiluminescence based tests. Materials and methods are provided for the chemical and/or physical control of conducting domains and reagent deposition for use in flat panel displays and multiply specific testing procedures.

Claims

1. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector and a support comprising an electrode capable of inducing electrochemiluminescence and a counterelectrode, said electrode having immobilized on a surface thereof a binding reagent capable of binding said analyte or a binding partner of said analyte.

4. An apparatus as recited in claims 1 or 2, wherein the immobilized reagent and electrode are substantially permissive to electron transfer between said electrode and an electrochemiluminescent label.

6. An apparatus as recited in claim 1 wherein said electrode is porous.

20. An apparatus as recited in claims 1, 7, 8, 11, wherein said electrode or said plurality of electrodes comprise a carbonaceous material.

41. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, further comprising an electrochemiluminescent moiety.

42. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, further comprising an electrochemiluminescent moiety containing an organometallic compound.

43. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, further comprising an electrochemiluminescent moiety containing a Ru-containing or Os-containing organometallic compound.

50. The apparatus of claims 1, 2, 7, 8 or 36, wherein said binding reagent is immobilized through hydrophobic interaction.

52. The apparatus of claims 1, 2, 7, 8 or 36, wherein said binding reagents are immobilized by covalent bonding.

54. The apparatus of claims 1, 2, 7, 8, 9 or 36, wherein said binding reagent comprises proteins or fragments or derivatives thereof.

67. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said light detector comprises film.

69. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said light detector comprises a photomultiplier.

70. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said electrochemiluminescence is imaged by using a photodiode.

71. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said electrochemiluminescence is imaged by using a photodiode array.

72. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said electrochemiluminescence is imaged by using a CCD array.

73. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, or 37, wherein said analyte is a nucleic acid.

74. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said electrochemiluminescence is imaged by using an array of light detectors.

75. The apparatus of claims 1, 9, 32, 33 or 35, wherein said support comprises a conductive layer and said conductive provides said electrode.

77. The apparatus of claims 1, 7, 8, 9, 21, 32, 33, 34 or 38, wherein said electrode comprises carbon nanotubes, carbon black, carbon nanotubes in a matrix, carbon black in a matrix or combinations thereof.

79. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34 or 38, wherein said electrode comprises carbon nanotubes.

81. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34 or 38, wherein said electrode comprises carbon nanotubes dispersed in a matrix.

88. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34 or 38, wherein said electrode comprises carbon particles dispersed in a matrix.

90. The apparatus of claims 1, 2, 7, 8, 9, 21, 32, 33, 34 or 38, wherein said electrode comprises functionalized carbon nanotubes.

91. The apparatus of claims 1 or 33, further comprising conductive projections from the surface of said support, said conductive projections capable of acting as said electrodes for generating electrochemiluminescence.

100. The apparatus of claims 1, 2, 7 or 8, wherein said binding reagent is present in a predetermined amount.

2. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector and an electrode capable of inducing electrochemiluminescence, said electrode having immobilized on a surface thereof a binding reagent capable of directly or indirectly binding said analyte or a binding partner of said analyte, wherein said electrode comprises carbon nanotubes, carbon black, carbon nanotubes in a matrix, carbon black in a matrix or combinations thereof.

3. An apparatus as recited in claim 2 wherein said apparatus further comprises a counterelectrode.

5. An apparatus as recited in claim 2 wherein said electrode is porous.

26. An article as recited in claim 5, wherein said porous material comprises carbon.

27. An article as recited in claim 5, which said porous material comprises fibrils.

101. The apparatus of claims 2, 7, 8, 9, 21, 32, 33 or 34, further comprising a counterelectrode in a fixed position relative to said electrode.

7. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector and a porous electrode capable of inducing electrochemiluminescence, said electrode having immobilized on a surface thereof a binding reagent capable of binding a component of a binding electrochemiluminescence assay.

8. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector and an electrode capable of inducing electrochemiluminescence and a porous matrix in electrochemical contact with said electrode or capable of being in electrochemical contact with said electrode, said porous matrix containing on a surface thereof a binding reagent capable of binding a component of a binding electrochemiluminescence assay.

9. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising:

(a) an electrode;

(b) a support having immobilized thereon a plurality of discrete binding domains each containing a binding reagent capable of binding a component of a binding electrochemiluminescence assay; and

(c) a light detector.

10. An apparatus as recited in claim 9 wherein said plurality of domains are in electrochemical contact with said electrode.

11. An apparatus as recited in claim 9 wherein said electrode is porous.

12. An apparatus as recited in claim 11 wherein said electrode is comprised of a carbonaceous material.

13. An apparatus as recited in claim 9 wherein said electrode is comprised of fibrils.

14. An apparatus as recited in claim 9 wherein said light detector detects electrochemiluminescence from two or more of said binding domains.

15. An apparatus as recited in claim 9 wherein said support is a porous matrix.

16. An apparatus as recited in claim 9 wherein said support is a porous matrix and said electrode is porous.

17. An apparatus as recited in claim 9 further comprising a counterelectrode, wherein said support is a porous matrix, said matrix being positioned between said electrode and said counterelectrode.

18. An apparatus as recited in claim 9, wherein said light detector comprises a plurality of light detectors and said plurality of light detectors is capable of imaging one or more of said binding domains.

19. An apparatus as recited in claim 18 wherein at least one of said plurality of light detectors detects electrochemiluminescence from one binding domain.

23. The apparatus of claim 9, said electrode and said discrete binding domains being in electrochemical contact.

24. An apparatus as recited in claim 9 wherein the electrode is the working electrode.

25. An apparatus as recited in claim 9 wherein the electrode is the counterelectrode.

21. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector and an electrode, said electrode having immobilized on a surface thereof a plurality of discrete binding domains, said binding domains being capable of binding a component of a binding electrochemiluminescence assay.

22. The apparatus of claim 21, wherein said discrete binding domains contain an electrochemiluminescent label.

44. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said plurality of discrete binding domains is in a range of from 5 through 1000 binding domains.

45. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said plurality of discrete binding domains is in a range of from 50 through 500 binding domains.

46. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said plurality of discrete binding domains is in a range of from 5 through 50 binding domains.

47. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said plurality of discrete binding domains includes greater than 50 binding domains.

48. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are in a size range of from 0.001 to 1 mm in diameter or width.

49. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are in the range of about 0.001 mm to about 0.01 mm in dimension.

51. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are immobilized through hydrophobic interaction.

53. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are attached by covalent bonding.

55. The apparatus of claims 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains include binding reagents comprising proteins or fragments or derivatives thereof.

56. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are located on a transparent gold film surface.

57. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains comprise patterned, self-assembled monolayers.

58. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains comprise patterned, self-assembled monolayers and said patterned self-assembled monolayers comprise alkane thiol.

61. The apparatus of claims 21, 33 or 38, wherein said binding domains are hydrophilic or hydrophobic relative to the surface of the electrode.

62. The apparatus according to claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, further comprising a domain that comprises an internal standard.

64. The apparatus of claims 21, 32, 33 or 34, wherein said binding domains and said electrode are substantially transparent.

68. The apparatus of claims 9, 21, 32, 33, 34, 35, 36, 37 or 39, wherein said light detector is capable of simultaneously measuring electrochemiluminescence from said binding domains and capable of distinguishing electrochemiluminescence emitted from different binding domains.

92. The apparatus of claims 9, 21, 32, 33, 34, 35, 37, 38 or 39, wherein said binding domains are contained in a single chamber capable of containing a single fluid in contact with two or more of said binding domains.

93. The apparatus of claims 7, 8, 21, 32, 33, 34 or 38, wherein said electrode is porous.

94. The apparatus of claims 7, 8, 21, 32, 33, 34 or 38, wherein said electrode is a flow-through electrode.

97. The apparatus of claims 7, 8, 9, 21, 32, 33, 34, 35, 36 or 37, further comprising a counterelectrode.

98. The apparatus of claims 9, 21, 32, 33, 35, 36, 37, 38 or 39, wherein one or more binding domains contain binding reagents of different specificity than those contained in other binding domains.

99. The apparatus of claims 9, 21, 32, 33, 34, 35, 36, 37, 38 or 39, wherein said binding domains comprise predetermined amounts of binding reagents.

28. A kit for use in performance of a plurality of electrochemiluminescence assays for analytes of interest comprising:

(a) an article comprising a plurality of discrete binding domains on a support, said binding domains having a relative spatial organization with respect to one another and at least two binding domains having different binding specificities for analytes of interest in an electrochemiluminescence assay; and

(b) a reagent comprising an electrochemiluminescent label.

29. A kit as recited in claim 28 further including an electrode.

103. The kit of claims 28 or 30, wherein said binding domains are contained in a single chamber capable of containing a single fluid in contact with two or more of said binding domains.

104. The kit of claims 28 or 30, wherein said binding domains are contained in a single chamber chamber capable of containing a single fluid in contact with two or more of said binding domains and said single chamber contains said reagent.

105. The kit of claim 28, wherein said support comprises a material selected from the group consisting of meshes, felts, carbon nanotubes, carbon black, porous materials, mats or mixtures thereof.

107. The kit of claim 28, wherein said support comprises:

(a) conducting polymers;

(b) conductive particles dispersed in a matrix;

(c) conductive fibers;

(d) a patterned self-assembled monolayer;

or

(e) mixtures thereof.

109. The kit of claims 28 or 30, wherein said electrochemiluminescent label comprises an organometallic compound.

110. The kit of claims 28 or 30, wherein said electrochemiluminescent label comprises an organometallic compound selected from the group consisting of Ru-containing and Os-containing organometallic compounds.

30. A kit for use in performance of a plurality of electrochemiluminescence assays for a plurality of analytes of interest comprising:

(a) an article comprising a plurality of discrete binding domains on an electrode, said binding domains having a relative spatial organization with respect to one another and at least two binding domains having different binding specificities for analytes of interest in an electrochemiluminescence assay; and

(b) a reagent comprising an electrochemiluminescent label.

106. The kit of claim 30, wherein said electrode comprises a material selected from the group consisting of meshes, felts, carbon nanotubes, carbon black, porous materials, mats or mixtures thereof.

108. The kit of claim 30, wherein said electrode comprises:

(a) conducting polymers;

(b) conductive particles dispersed in a matrix;

(c) conductive fibers;

(d) a patterned self-assembled monolayer;

or

(e) mixtures thereof.

31. A system for conducting a plurality of electrochemiluminescence assays comprising:

(a) a plurality of discrete binding domains specific for a plurality of different analytes;

(b) a voltage waveform generator; and

(c) a light detector capable of simultaneously detecting light from more than one binding domain.

32. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising:

(a) a support having a plurality of discrete binding domains thereon;

(b) an electrode capable of inducing electrochemiluminescence, wherein said plurality of discrete binding domains and said electrode are capable of being brought into proximity;

(c) a source of electrical energy for generating electrochemiluminescence; and

(d) a light detector.

40. The apparatus of claims 9, 32, 33 or 34, wherein said support further comprises a counterelectrode.

59. The apparatus of claims 9, 32, 33, 34, or 35, wherein said binding domains are formed by a method comprising the steps of:

(a) linking a group A to at least one binding domain on said support, said group A capable of specifically binding to a second group B; and

(b) contacting said group A with said group B, said group B being bound to, associated with or integral to a binding reagent for an analyte of interest, thereby forming a binding surface containing said binding reagent linked to said support via an A;

B linkage, said binding surface being organized as said plurality of discrete binding domains containing said binding reagents capable of binding to an analyte of interest.

60. The apparatus of claims 9, 32, 33, 34 or 35, wherein said binding domains are hydrophilic or hydrophobic relative to the surface of the support.

63. The apparatus of claims 32, 35, 37, 38 or 39, wherein said support, binding domains and electrodes are substantially transparent.

65. The apparatus of claims 32 or 33, wherein said binding domains and said support are substantially transparent.

76. The apparatus of claims 9, 32, 33 or 35, wherein said support comprises meshes, felts, fibrous materials, gels, elastomers or mixtures thereof.

83. The apparatus of claims 9, 32, 33 or 35, wherein said support comprises graphite, glassy carbon or carbon black.

84. The apparatus of claims 9, 32, 33 or 35, wherein said support comprises graphitic carbon nanotubes.

85. The apparatus of claims 9, 32, 33 or 35, wherein said support comprises carbon particles dispersed in a matrix.

86. The apparatus of claims 32, 33, 34 or 38, wherein said electrode comprises graphite, glassy carbon or carbon black.

33. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising:

(a) a support having a plurality of discrete binding domains thereon, wherein said support comprises an electrode for generating electrochemiluminescence;

(b) a source of electrical energy for generating electrochemiluminescence; and

(c) a light detector.

34. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising:

(a) a cassette, wherein said cassette comprises an electrode and a plurality of discrete binding domains, wherein one or more binding domains contain binding reagents of specificity different from other binding domains;

(b) a receptacle for said cassette;

(c) a source of electrical energy for generating electrochemiluminescence; and

(d) a light detector.

35. A apparatus for use in the detection of an analyte by electrochemiluminescence comprising:

(a) a plurality of discrete binding domains immobilized on a support; and

(b) a plurality of electrodes capable of inducing electrochemiluminescence, wherein said binding domains are spatially aligned with said plurality of electrodes.

66. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes are separately addressable by at least one source of electrical energy.

78. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes comprise carbon nanotubes, carbon black, carbon nanotubes in a matrix, carbon black in a matrix or combinations thereof.

80. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes comprise carbon nanotubes.

82. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes comprise carbon nanotubes dispersed in a matrix.

87. The apparatus of claims 35, 36, 37 or 39, wherein said plurality of electrodes comprise graphite, glassy carbon or carbon black.

89. The apparatus of claims 35, 36, 37 or 38, wherein said electrodes comprise carbon particles dispersed in a matrix.

95. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes are porous.

96. The apparatus of claims 35, 36, 37 or 39, wherein said electrodes are flow-through electrodes.

102. The apparatus of claims 35, 36, 37 or 38, further comprising a plurality of counterelectrodes to form a plurality of electrode-counterelectrode pairs.

36. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a plurality of electrodes capable of inducing electrochemiluminescence and a light detector, said electrodes having immobilized on surfaces thereof binding reagents capable of binding components of a binding electrochemiluminescence assay.

37. An apparatus for use in the detection of an analyte by electrochemiluminescence comprising a light detector, a plurality of electrodes capable of inducing electrochemiluminescence and a plurality of discrete binding domains proximate said electrodes, said plurality of electrodes being capable of inducing electrochemiluminescence from at least one binding domain, each binding domain having an amount of binding reagent capable of binding a component of a binding electrochemiluminescence assay.

38. An apparatus for generating electrochemiluminescence comprising:

(a) a support;

(b) an electrode on said support having a plurality of discrete binding domains immobilized thereon;

(c) a counterelectrode on said support; and

(d) a light detector.

39. An apparatus for generating electrochemiluminesence comprising:

(a) a support;

(b) a plurality of electrodes on said support, said electrodes having binding domains immobilized thereon;

(c) a counterelectrode on said support; and

(d) a light detector.

111. A kit for use in performance of a plurality of electrochemiluminescence assays for a plurality of analytes of interest comprising:

(a) an article comprising a plurality of discrete binding domains on a support, said binding domains having a relative spatial organization with respect to one another and having different binding specificities for a plurality of different analytes of interest in an electrochemiluminescence assay; and

(b) a vessel containing a reagent necessary for conducting said assays.

112. A kit as recited in claim 111, further including an apparatus for performing said assays.

116. The kit as recited in claim 111, wherein said plurality of binding domains includes binding domains having different binding specificities to provide for simultaneous binding of a plurality of different analytes of interest present in a sample.

117. The kit as recited in claim 111, wherein said support comprises a surface which is selectively derivatized to comprise one or more specific binding components.

118. The kit as recited in claim 117, wherein said surface comprises a self-assembling monolayer.

119. The kit as recited in claim 117, wherein said surface is a matrix selected from the group consisting of a conductor, a metal electrode, a conducting polymer film, an insulator, a semi-conductor, an ionic conductor, and a porous material.

113. A kit for use in performance of a plurality of electrochemiluminescence assays for a plurality of analytes of interest comprising:

a vessel containing binding components specific for a plurality of different analytes for use in a plurality of electrochemiluminescence assays, wherein at least two of said binding components have specificity for different analytes and being linked to electrochemiluminescent labels having wavelengths of emission that are substantially identical.

114. A kit as recited in claim 113, further containing one or more vessels containing non-binding components for a plurality of electrochemiluminescence assays.

115. A kit as recited in claim 113, further containing an article comprising a plurality of discrete binding domains on a support, said binding domains having a relative spatial organization with respect to one another and having different binding specificities for a plurality of different analytes of interest in an electrochemiluminescence assay.