Electronic detection of biological molecules using thin layers

• US 6,824,974 B2
• Filed: 10/02/2001
• Issued: 11/30/2004
• Est. Priority Date: 06/11/2001
• Status: Expired due to Term

• Current Assignee: Bridger Technologies, Inc. (Bio-Rad Laboratories Incorporated)
• Original Assignee: Genorx, Inc.
• Inventors: Kunwar, Sandeep, Pisharody, Sobha M., Mathai, George T.
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US09/970,087
• Publication Number:
  US 20040146863A1
• Time in Patent Office: 1,155 Days
• Field of Search: 435/6, 435/91.2, 205/777.5, 536/23.1
• US Class Current: 435/4
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/116 electrical properties of nucleic acids, e.g. impedance, conductivity or resistance
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G11C 13/0014 comprising cells based on organic memory material
• CPC Class Codes: G11C 13/0019 comprising bio-molecules
• CPC Class Codes: Y10S 977/957 Of chemical property or presence

Abstract

This invention provides novel sensors that facilitate the detection of essentially any analyte. In general, the biosensors of this invention utilize a binding agent (e.g. biomolecule) to specifically bind to one or more target analytes. In preferred embodiments, the biomolecules spans a gap between two electrodes. Binding of the target analyte changes conductivity of the sensor thereby facilitating ready detection of the binding event and thus detection and/or quantitation of the bound analyte. A molecular sensing apparatus comprising.

Claims

1. A molecular sensing apparatus comprising:

a plurality of electrode pairs, wherein the electrode pairs comprise;

a first electrode;

a second electrode; and

an insulator between said first electrode and said second electrode, wherein said insulator comprises a channel between said first electrode and said second electrode and wherein said first electrode and said second electrode are separated by less than 30 nanometers

and said apparatus is configured by having parallel electrode portions to permit the formation of a plurality of independent, electrically coupled binding agent/analyte complexes electrically in parallel between said first electrode and said second electrode.

2. The molecular sensing apparatus of claim 1, wherein said insulator has a resistivity greater than 10⁻

3 ohm-meters.

3. The molecular sensing apparatus of claim 1, wherein said insulator is selected from the group consisting of SiO₂, TiO₂, ZrO₂, quartz, porcelain, ceramic, polystyrene, TEFLON, and an insulating oxide or sulfide of a transition metal in the periodic table of the elements.

4. The molecular sensing apparatus of claim 1, wherein said first electrode and said second electrode are separated by a distance in the range of 1 Angstrom to 10¹⁰ Angstroms.

5. The molecular sensing apparatus of claim 1, wherein at least one of said first electrode and said second electrode has a resistivity of less than 10⁻

2 ohm-meters.

6. The molecular sensing apparatus of claim 1, wherein at least one of said first electrode and said second electrode has a resistivity of less than 10⁻

3 ohm-meters.

7. The molecular sensing apparatus of claim 1, wherein said first electrode and said second electrode each comprises a material selected from the group consisting of ruthenium, osmium, cobalt, rhodium, rubidium, lithium, sodium, potassium, vanadium, cesium, beryllium, magnesium, calcium, chromium, molybdenum, silicon, germanium, aluminum, iridium, nickel, palladium, platinum, iron, copper, titanium, tungsten, silver, gold, zinc, cadmium, indium tin oxide, carbon, and carbon nanotube.

8. The molecular sensing apparatus of claim 1, wherein at least one of said first electrode and said second electrode is functionalized with a chemical group that can be derivatized or crosslinked.

9. The molecular sensing apparatus of claim 8, wherein said chemical group is selected from the group consisting of a sulfate, a sulfhydryl, an amine, an aldehyde, a carboxylic acid, a phosphate, a phosphonate, an alkene, an alkyne, a hydroxyl, a bromine, an iodine, a chlorine, a light-activatable group, and a group activatable by an electric potential.

10. The molecular sensing apparatus of claim 1, wherein at least one of said first electrode and said second electrode bears a self-assembled monolayer (SAM).

11. The molecular sensing apparatus of claim 10, wherein said SAM comprises a compound selected from the group consisting of an alkanethiol, a phospholipid, a bola amphiphile, and an oligo (phenylenevinylene).

12. The molecular sensing apparatus of claim 1, further comprising a substrate that supports the first electrode and the second electrode, wherein the first electrode and the second electrode are integrated with the substrate.

13. The molecular sensing apparatus of claim 1, wherein the first electrode and the second electrode are integrated with the insulator to form a substrate.

14. The molecular sensing apparatus of claim 1, wherein said first electrode comprises a surface with a shape selected from the group consisting of convex, concave, textured, corrugated, patterned uniformly, and randomly patterned.

15. The molecular sensing apparatus of claim 1, wherein said first electrode and said second electrode are oriented in a formation selected from the group consisting of annular, planar, and orthogonal.

16. The molecular sensing apparatus of claim 1, wherein the first electrode has a first surface and a said second electrode has a second surface, wherein the first surface is not coplanar to the second surface.

17. The apparatus of claim 1, wherein said at least one electrode pair comprises a first electrode pair and a second electrode pair.

18. The molecular sensing apparatus of claim 1, wherein said one or more electrode pairs are at least 10 electrode pairs.

19. The molecular sensing apparatus of claim 1, wherein said one or more electrode pairs are at least 1,000 electrode pairs.

20. The molecular sensing apparatus of claim 1, wherein said one or more electrode pairs comprises about 10² to 10¹⁰ electrode pairs.

21. The molecular sensing apparatus of claim 1, the molecular sensing apparatus further comprising a measurement device electrically coupled to each first electrode and to each second electrode of each electrode pair in said at least one electrode pair.

22. The molecular sensing apparatus of claim 21, wherein said measurement device measures an electromagnetic property selected from the group consisting of direct electric current, alternating electric current, permitivity, resistivity, electron transfer, electron tunneling, electron hopping, electron transport, electron conductance, voltage, electrical impedance, signal loss, dissipation factor, resistance, capacitance, inductance, magnetic field, electrical potential, charge and magnetic potential.

23. The molecular sensing apparatus of claim 1, further comprising an electrical circuit electrically coupled to the first electrode and the second electrode.

24. The molecular sensing apparatus of claim 23, wherein said electrical circuit comprises an electric signal gating system.

25. The molecular sensing apparatus of claim 24, wherein said electric signal gating system comprises a CMOS gating system.

26. The apparatus of claim 17, wherein

a first biological macromolecule is attached to the first electrode and the second electrode in the first electrode pair, and

a second biological macromolecule is attached to the first electrode and the second electrode in the second electrode pair; wherein the first biological molecule and the second biological molecule are the same.

27. The apparatus of claim 17, wherein

a first biological macromolecule is attached to the first electrode and the second electrode in the first electrode pair, and

a second biological macromolecule is attached to the first electrode and the second electrode in the second electrode pair;

wherein the first biological molecule and the second biological molecule are different.

28. The molecular sensing apparatus of claim 1, further comprising a computer electrically coupled to the first electrode and the second electrode.

29. The molecular sensing apparatus of claim 1, wherein at least one of the first electrode and the second electrode comprises a semiconductor material.

30. The molecular sensing apparatus of claim 29, wherein said semiconductor material has a resistivity ranging from 10⁻

6ω

-m to 10⁷Ω

-m.

31. The molecular sensing apparatus of claim 29, wherein the semiconductor material is selected from the group consisting of silicon, dense silicon carbide, boron carbide, Fe₃O₄, germanium, silicon germanium, silicon carbide, tungsten carbide, titanium carbide, indium phosphide, gallium nitride, gallium phosphide, aluminum phosphide, aluminum arsenide, mercury cadmium telluride, tellurium, selenium, ZnS, ZnO, ZnSc. CdS, ZnTc, GaSc, CdSe, CdTe, GaAs, InP, GaSb, EnAs, Te, PbS, InSb, PbTe, PbSe, and tungsten disulfide.

60. The molecular sensing apparatus of claim 1, wherein a biological macromolecule or macromolecule/analyte complex connects said first electrode and said second electrode in said first electrode pair.

61. The molecular sensing apparatus of claim 60 wherein said biological macromolecule is selected from the group consisting of a nucleic acid, a protein, a polysaccharide, a lectin, and a sugar.

62. The molecular sending apparatus of claim 60 wherein said biological macromolecule is a deoxyribonucleic acid or a nucleic acid.

63. The molecular sensing apparatus of claim 60 wherein said biological macromolecule is functionalized with a chemical group selected from the group consisting of a sulfate, a sulfhydryl, an amine, an aldehyde, a carboxylic acid, a phosphate, a phosphonate, an alkene, an alkyne, a hydroxyl, a bromine, an iodine, a chlorine, a light-activatable group, and a group activatable by an electric potential.

64. The molecular sensing apparatus of claim 60 wherein the biological macromolecule is attached to the first electrode by a thiol group.

65. The molecular sensing apparatus of claim 60 wherein the biological macromolecule is attached to the first electrode by a phosphorothioate or a phosphonate.

66. The molecular sensing apparatus of claim 60 wherein the biological macromolecule is attached to said first electrode by a linker.

67. The molecular sensing apparatus of claim 66 wherein said linker is selected from the group consisting of DFDNB, DST, ABH, ANB-NOS, EDC, NHS-ASA, and SIA.

68. The molecular sensing apparatus of claim 1 wherein a first biological macromolecule is attached to said first electrode and a second biological macromolecule is attached to said second electrode.

69. The molecular sensing apparatus of claim 1 wherein said first electrode comprises a surface with a shape selected from the group consisting of convex, concave, textured, corrugated, patterned uniformly, and randomly patterned.

70. The molecular sensing apparatus of claim 62 wherein said nucleic acid is deoxyribonucleic acid or ribonucleic acid.

71. The molecular sensing apparatus of claim 1, wherein said first electrode and said second electrode are separated by less than 20 nanometers.

72. The molecular sensing apparatus of claim 71, wherein said first electrode and said second electrode are separated by less than 15 nanometers.

73. The molecular sensing apparatus of claim 72, wherein said first electrode and said second electrode are separated by less than 10 nanometers.

32. A molecular sensing apparatus comprising a plurality of electrode pairs in an insulating substrate,

wherein a first electrode pair in said plurality of electrode pairs comprises a first electrode and a second electrode,

wherein said first electrode and said second electrode are separated by less than 30 nanometers and said apparatus is configured by having parallel electrode portions to permit the formation of a plurality of independent, electrically coupled binding agent/analyte complexes electrically in parallel between said first electrode and said second electrode.

33. The molecular sensing apparatus of claim 32 wherein a biological macromolecule connects said first electrode and said second electrode.

34. The molecular sensing apparatus of claim 33 wherein said biological macromolecule is a nucleic acid.

35. The molecular sensing apparatus of claim 34 wherein said nucleic acid is a deoxyribonucleic acid or a ribonucleic acid.

36. The molecular sensing apparatus of claim 32 wherein at least one of said first electrode and said second electrode has a resistivity of less than 10⁻

3 ohm-meters.

37. The molecular sensing apparatus of claim 32 wherein said first electrode and said second electrode comprise a material selected from the group consisting of ruthenium, osmium, cobalt, rhodium, rubidium, lithium, sodium, potassium, vanadium, cesium, beryllium, magnesium, calcium, chromium, molybdenum, silicon, germanium, aluminum, iridium, nickel, palladium, platinum, iron, copper, titanium, tungsten, silver, gold, zinc, cadmium, indium tin oxide, carbon, and carbon nanotube.

38. The molecular sensing apparatus of claim 32 wherein at least one of said first electrode and said second electrode is functionalized with a chemical group that can be derivatized or crosslinked.

39. The molecular sensing apparatus of claim 38 wherein said chemical group is a sulfate, a sulfhydryl, an amine, an aldehyde, a carboxylic acid, a phosphate, a phosphonate, an alkene, an alkyne, a hydroxyl, a bromine, an iodine, a chlorine, a light-activatable group, or a group activatable by an electric potential.

40. The molecular sensing apparatus of claim 32 wherein at least one of said first electrode and said second electrode is coated with a self-assembled monolayer.

41. The molecular sensing apparatus of claim 40 wherein said self-assembled monolayer comprises a compound selected from the group consisting of an alkanethiol, a phospholipid, a bola amphiphile, and an oligo(phenylenevinylene).

42. The molecular sensing apparatus of claim 33 wherein the biological macromolecule is attached to the first electrode by a thiol group.

43. The molecular sensing apparatus of claim 34 wherein the biological macromolecule is attached to the first electrode by a phosphonate.

44. The molecular sensing apparatus of claim 34 wherein the biological macromolecule is attached to said first electrode by a linker.

45. The molecular sensing apparatus of claim 44 wherein said linker is selected from the group consisting of DFDNB, DST, ABH, ANB-NOS, EDC, NHS-ASA, and SIA.

46. The molecular sensing apparatus of claim 32 wherein the first electrode has a first surface and the second electrode has a second surface, and wherein the first surface is not coplanar to the second surface.

47. The molecular sensing apparatus of claim 32 wherein said one or more electrode pairs comprise at least three electrode pairs.

48. The molecular sensing apparatus of claim 32 wherein said one or more electrode pairs comprise at least 10,000 electrode pairs.

49. The molecular sensing apparatus of claim 32 wherein said one or more electrode pairs comprises 10² to 10¹⁰ electrode pairs.

50. The molecular sensing apparatus of claim 32 the apparatus further comprising a measurement device electrically coupled to the first electrode and to the second electrode said first electrode pair.

51. The molecular sensing apparatus of claim 50 wherein said measurement device measures an electromagnetic property selected from the group consisting of direct electric current, alternating electric current, permitivity, resistivity, electron transfer, electron tunneling, electron hopping, electron transport, electron conductance, voltage, electrical impedance, signal loss, dissipation factor, resistance, capacitance, inductance, magnetic field, electrical potential, charge and magnetic potential.

52. The molecular sensing apparatus of claim 32 further comprising an electrical circuit electrically coupled to the first electrode and the second electrode of said first electrode pair.

53. The molecular sensing apparatus of claim 52 wherein said electrical circuit comprises an electric signal gating system.

54. The molecular sensing apparatus of claim 32 wherein said biological molecule connects to said first electrode and said second electrode in said first electrode pair.

55. The molecular sensing apparatus of claim 32 wherein

a first biological macromolecule is attached to said first electrode in said first electrode pair, and

a second biological macromolecule is attached to said second electrode in said first electrode pair.

56. The molecular sensing apparatus of claim 32 further comprising a computer electrically coupled to the first electrode and the second electrode of at least one electrode pair in said one or more electrode pairs.

57. The molecular sensing apparatus of claim 32 wherein at least one of the first electrode and the second electrode in an electrode pair in said one or more electrode pairs comprises a semiconductor material.

58. The molecular sensing apparatus of claim 57 wherein said semiconductor material has a resistivity between 10⁻

6Ω

-m and 10<sup>−

7</sup>Ω

-m.

59. The molecular sensing apparatus of claim 57 wherein the semiconductor material is selected from the group, consisting of silicon, dense silicon carbide, boron carbide, Fe₃O₄, germanium, silicon germanium, silicon carbide, tungsten carbide, titanium carbide, indium phosphide, gallium nitride, gallium phosphide, aluminum phosphide, aluminum arsenide, mercury cadmium telluride, tellurium, selenium, ZnS, ZnO, ZnSe, CdS, ZnTe, GaSe, CdSe, CdTe, GaAs, InP, GaSb, EnAs, Te, PbS, InSb, PbTe, PbSe, and tungsten disulfide.

74. The molecular sensing apparatus of claim 32, wherein said first electrode and said second electrode are separated by less than 20 nanometers.

75. The molecular sensing apparatus of claim 74, wherein said first electrode and said second electrode are separated by less than 15 nanometers.

76. The molecular sensing apparatus of claim 75, wherein said first electrode and said second electrode are separated by less than 10 nanometers.