Mediator-modified redox biomolecules for use in electrochemical determination of analyte

• US 20060025593A1
• Filed: 07/28/2004
• Published: 02/02/2006
• Est. Priority Date: 07/28/2004
• Status: Active Grant

• Current Assignee: Agency For Science Technology and Research (Government of Singapore)
• Original Assignee: Agency For Science Technology and Research (Government of Singapore)
• Inventors: Gao, Zhiqiang, Xie, Hong
• Granted Patent: US 7,511,142 B2
• Time in Patent Office: Days
• US Class Current: 546/2
• CPC Class Codes: C07F 15/0026 without a metal-carbon linkage

Abstract

Compositions and methods for electrochemical detection of an analyte comprising a transition metal compound wherein M is a metallic element that can form a coordinate bond to nitrogen; R and R′ are coordinated to M at their nitrogen atoms; L is a linking ligand; Z is chlorine or bromine; m can be from 1 to 6 and X is an anion, or combination of anions, that balances the charge m. Also provided are electrochemical tags and methods of detection.

Claims

1. A transition metal compound of the formula:

2. A compound as set forth in claim 1 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

3. A compound as set forth in claim 2 wherein M is osmium.

4. A compound as set forth in claim 1 wherein the linking ligand comprises 3-aminopropylimidazole.

5. A compound as set forth in claim 1 wherein Z is chlorine or bromine.

6. A compound as set forth in claim 1 wherein R and R′

are independently selected from the group consisting of 2,2′

-bipyridyl, 4,4′

-methyl-2,2′

-bipyridyl, 4,4′

-ethyl-2,2′

-bipyridyl, 4,4′

-phenyl-2,2′

-bipyridyl, 5,5′

-methyl-2,2′

-bipyridyl, 5,5′

-ethyl-2,2′

-bipyridyl and 5,5′

-phenyl-2,2′

-bipyridyl.

7. A compound as set forth in claim 6 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

8. A compound as set forth in claim 7 wherein M is osmium.

9. A compound as set forth in claim 6 wherein the linking ligand comprises 3-aminopropylimidazole.

10. A compound as set forth in claim 6 wherein Z is chlorine or bromine.

11. A compound as set forth in claim 1 wherein at least one of R and R′

is 2,2′

-bipyridyl, M is osmium, Z is chlorine and the linking ligand comprises 3-aminopropylimidazole.

12. A compound as set forth in claim 1 comprising the formula:

13. A method for preparing a transition metal compound, the method comprising contacting a precursor compound of the formula:

14. A method as set forth in claim 13 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

15. A method as set forth in claim 14 wherein M is osmium.

16. A method as set forth in claim 13 wherein the linking ligand comprises 3-aminopropylimidazole.

17. A method as set forth in claim 13 wherein Z is chlorine or bromine.

18. A method as set forth in claim 13 wherein R and R′

are independently selected from the group consisting of 2,2′

-bipyridyl, 4,4′

-methyl-2,2′

-bipyridyl, 4,4′

-ethyl-2,2′

-bipyridyl, 4,4′

-phenyl-2,2′

-bipyridyl, 5,5′

-methyl-2,2′

-bipyridyl, 5,5′

-ethyl-2,2′

-bipyridyl and 5,5′

-phenyl-2,2′

-bipyridyl.

19. A method as set forth in claim 18 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

20. A method as set forth in claim 19 wherein M is osmium.

21. A method as set forth in claim 18 wherein the linking ligand comprises 3-aminopropylimidazole.

22. A method as set forth in claim 18 wherein Z is chlorine or bromine.

23. A method as set forth in claim 13 wherein at least one of R and R′

is 2,2′

-bipyridyl, M is osmium, Z is chlorine and the linking ligand comprises 3-aminopropylimidazole.

24. A method as set forth in claim 13, wherein the precursor compound is a compound of the formula:

25. An electrochemical tag comprising a redox biomolecule bonded to a transition metal compound of the formula:

26. An electrochemical tag as set forth in claim 25 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

27. An electrochemical tag as set forth in claim 26 wherein M is osmium.

28. An electrochemical tag as set forth in claim 25 wherein the redox biomolecule comprises an oxidoreductase.

29. An electrochemical tag as set forth in claim 28 wherein the oxidoreductase comprises glucose oxidase.

30. An electrochemical tag as set forth in claim 25 wherein the linking ligand comprises 3-aminopropylimidazole.

31. An electrochemical tag as set forth in claim 25 wherein Z is chlorine or bromine.

32. An electrochemical tag as set forth in claim 25 wherein R and R′

are independently selected from the group consisting of 2,2′

-bipyridyl, 4,4′

-methyl-2,2′

-bipyridyl, 4,4′

-ethyl-2,2′

-bipyridyl, 4,4′

-phenyl-2,2′

-bipyridyl, 5,5′

-methyl-2,2′

-bipyridyl, 5,5′

-ethyl-2,2′

-bipyridyl and 5,5′

-phenyl-2,2′

-bipyridyl.

33. An electrochemical tag as set forth in claim 32 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

34. An electrochemical tag as set forth in claim 33 wherein M is osmium.

35. An electrochemical tag as set forth in claim 32 wherein the redox biomolecule comprises an oxidoreductase.

36. An electrochemical tag as set forth in claim 35 wherein the oxidoreductase comprises glucose oxidase.

37. An electrochemical tag as set forth in claim 32 wherein the linking ligand comprises 3-aminopropylimidazole.

38. An electrochemical tag as set forth in claim 32 wherein Z is chlorine or bromine.

39. An electrochemical tag as set forth in claim 25 wherein at least one of R and R′

is 2,2′

-bipyridyl, M is osmium, Z is chlorine and the linking ligand comprises 3-aminopropylimidazole.

40. An electrochemical tag as set forth in claim 39 wherein the redox biomolecule comprises an oxidoreductase.

41. An electrochemical tag as set forth in claim 40 wherein the oxidoreductase comprises glucose oxidase.

42. An electrochemical tag as set forth in claim 25 comprising a redox biomolecule bonded to a transition metal compound of the formula:

43. An electrochemical tag as set forth in claim 42 wherein the redox biomolecule comprises an oxidoreductase.

44. An electrochemical tag as set forth in claim 43 wherein the oxidoreductase comprises glucose oxidase.

45. A method for preparing an electrochemical tag as set forth in claim 25, the method comprising bonding a redox biomolecule to a transition metal compound of the formula:

46. A method as set forth in claim 45 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

47. A method as set forth in claim 46 wherein M is osmium.

48. A method as set forth in claim 45 wherein the redox biomolecule comprises an oxidoreductase.

49. A method as set forth in claim 48 wherein the oxidoreductase comprises glucose oxidase.

50. A method as set forth in claim 45 wherein the redox biomolecule is bound to the transition metal compound by an amide bond.

51. A method as set forth in claim 45 wherein the linking ligand comprises 3-aminopropylimidazole.

52. A method as set forth in claim 45 wherein Z is chlorine or bromine.

53. A method as set forth in claim 45 wherein R and R′

are independently selected from the group consisting of 2,2′

-bipyridyl, 4,4′

-methyl-2,2′

-bipyridyl, 4,4′

-ethyl-2,2′

-bipyridyl, 4,4′

-phenyl-2,2′

-bipyridyl, 5,5′

-methyl-2,2′

-bipyridyl, 5,5′

-ethyl-2,2′

-bipyridyl and 5,5′

-phenyl-2,2′

-bipyridyl.

54. A method as set forth in claim 53 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

55. A method as set forth in claim 54 wherein M is osmium.

56. A method as set forth in claim 53 wherein the redox biomolecule comprises an oxidoreductase.

57. A method as set forth in claim 56 wherein the oxidoreductase comprises glucose oxidase.

58. A method as set forth in claim 53 wherein the redox biomolecule is bound to the transition metal compound by an amide bond.

59. A method as set forth in claim 53 wherein the linking ligand comprises 3-aminopropylimidazole.

60. A method as set forth in claim 53 wherein Z is chlorine or bromine.

61. A method as set forth in claim 45 wherein at least one of R and R′

is 2,2′

-bipyridyl, M is osmium, Z is chlorine and the linking ligand comprises 3-aminopropylimidazole.

62. A method as set forth in claim 61 wherein the redox biomolecule comprises an oxidoreductase.

63. A method as set forth in claim 62 wherein the oxidoreductase comprises glucose oxidase.

64. A method as set forth in claim 61 wherein the redox biomolecule is bound to the transition metal compound by an amide bond.

65. A method as set forth in claim 45, the method comprises binding a redox biomolecule to a transition metal compound of the formula:

66. A method as set forth in claim 65 wherein the redox biomolecule comprises an oxidoreductase.

67. A method as set forth in claim 66 wherein the oxidoreductase comprises glucose oxidase.

68. A method as set forth in claim 66 wherein the redox biomolecule is bound to the transition metal compound by an amide bond.

69. An electrochemical method for detecting an analyte in a sample, the method comprising:

forming at an electrode surface an electrochemically active redox complex comprising the analyte and an electrochemical tag; and

detecting electron transfer at the electrode, wherein the electrochemical tag comprises a redox biomolecule bonded to a transition metal compound of the formula;

wherein

M is a metallic element that can form a coordinate bond to nitrogen;

R and R′

are nitrogen-containing organic moieties coordinated to M at their nitrogen atoms, wherein

R and R′

are independently selected from the group consisting of 2,2′

-bipyridyl;

2,2′

-bipyridyl substituted with one or more substituents selected from the group consisting of C1-C4 alkyl, phenyl and phenyl substituted with one or more C1-C4 alkyl groups;

1,10-phenanthrolinyl; and

1,10-phenanthrolinyl substituted with one or more substituents selected from the group consisting of C1-C4 alkyl, phenyl and phenyl substituted with one or more C1-C4 alkyl groups;

L is a linking ligand comprising an organic amine having from about 3 to about 20 non-hydrogen atoms, an aliphatic amino group and a nitrogenous moiety that provides a metal-to-nitrogen coordinate bond to M;

Z is a halogen atom;

m is +1, +2, +3, +4, +5, or +6; and

X is an anion, or combination of anions, that balances m.

70. A method as set forth in claim 69 wherein M is selected from the group consisting of osmium, ruthenium, zinc, iron, rhodium, rhenium, platinum, scandium, titanium, vanadium, cadmium, magnesium, copper, cobalt, palladium, chromium, manganese, nickel, molybdenum, tungsten and iridium or mixtures thereof.

71. A method as set forth in claim 70 wherein M is osmium.

72. A method as set forth in claim 69 wherein the analyte is an oxidation substrate for the redox biomolecule.

73. A method as set forth in claim 69 wherein the redox biomolecule comprises an oxidoreductase.

74. A method as set forth in claim 73 wherein the oxidoreductase comprises glucose oxidase.

75. A method as set forth in claim 69 wherein the linking ligand comprises 3-aminopropylimidazole.

76. A method as set forth in claim 69 wherein Z is chlorine or bromine.

77. A method as set forth in claim 69, wherein the method further comprises:

immobilizing a probe onto the electrode surface;

labeling the analyte with a first member of a recognition pair;

labeling the electrochemical tag with a second member of the recognition pair; and

combining the labeled components with an oxidation substrate of the redox biomolecule in solution in contact with the electrode, wherein the probe and analyte specifically bind or hybridize to each other.

78. A method as set forth in claim 77, wherein the recognition pair comprises biotin and avidin.

79. A method as set forth in claim 77, wherein the analyte comprises a nucleic acid.

80. A method as set forth in claim 69, wherein the method further comprises:

labeling the electrode surface with a first member of a recognition pair;

labeling the electrochemical tag with a second member of the recognition pair; and

combining the labeled components with an oxidation substrate of the redox biomolecule in solution in contact with the electrode, wherein the electrochemical tag binds to the electrode surface.

81. A method as set forth in claim 80, wherein the recognition pair comprises biotin and avidin.

82. A method as set forth in claim 80, wherein the analyte comprises the substrate of the redox biomolecule.

83. A method as set forth in claim 69, wherein the transition metal compound comprises a +3 cation having the structure

84. A method as set forth in claim 83 wherein the redox biomolecule comprises an oxidoreductase.

85. A method as set forth in claim 83 wherein the oxidoreductase comprises glucose oxidase.