Detection of analytes using reorganization energy

• US 6,013,459 A
• Filed: 06/12/1997
• Issued: 01/11/2000
• Est. Priority Date: 06/12/1997
• Status: Expired due to Term

• Current Assignee: Osmetech Technology Inc. (Roche Holding AG)
• Original Assignee: Clinical Micro Sensors Inc. (Roche Holding AG)
• Inventors: Meade, Thomas J.
• Primary Examiner(s): Achutamurthy, Ponnathapura
• Assistant Examiner(s): Monshipouri, Maryam
• Application Number:
  US08/873,977
• Time in Patent Office: 943 Days
• Field of Search: 435/4, 435/5, 435/6, 435/7.1, 435/7.2, 435/7.3, 435/7.4-7.7, 435/7.71-7.72, 435/7.8-7.92, 205/77.5, 205/777.5, 205/778, 205/779, 205/793.5, 422/68.1, 422/82.01, 422/82.02
• US Class Current: 435/7.1
• 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: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

The invention relates to novel methods and compositions for the detection of analytes using the nuclear reorganization energy, λ, of an electron transfer process.

Claims

1. A method of detection a target analyte in a test sample comprising:

a) adding an analyte to an electrode comprising;

i) a solvent accessible transition metal complex comparing a metal selected from the group consisting of manganese, technetium, rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel, palladium, platinum, copper, silver and gold; and

ii) a binding ligand that will bind the target analyte;

such that upon binding of said analyte to said binding ligand, a solvent inhibited transition metal complex is formed; and

b) detecting electron transfer between said solvent inhibited transition metal complex and said electrode.

2. A method according to claim 1, wherein said solvent accessible transition metal complex has a solvent reorganization energy of greater than about 1200 mV and said solvent inhibited transition metal complex has a solvent reorganization energy of less than 1000 mV.

3. A method according to claim 1, wherein the solvent reorganization energy of said solvent inhibited transition metal complex decreases by at least 100 mV upon binding of said analyte to form said solvent inhibited transition metal complex.

4. A method according to claim 1, wherein upon binding, at least one solvent accessible transition metal complex is less than 8 Å

from the bound analyte such that it forms said solvent inhibited transition metal complex.

5. A method according to claim 1, wherein said solvent accessible transition metal complex has at least one coordination site occupied a polar coordination group.

6. A method according to claim 1 further comprising applying at least a first input signal to said solvent inhibited transition metal complex.

7. A method according to claim 6 wherein in the absence of target analyte, said first input signal does not result in significant electron transfer.

8. A method according to claim 6, wherein said first input signal comprises at least an AC component.

9. A method according to claim 6 further comprising applying input signal at a plurality of frequencies.

10. A method according to claim 6 or 8, wherein said first input signal comprises at least a DC component.

11. A method according to claim 10 further comprising applying input signal at a plurality of voltages.

12. A method according to claim 1 wherein said detecting is by receiving an output signal characteristic of electron transfer between said solvent inhibited transition metal complex and said electrode.

13. A method according to claim 12 wherein said output signal is a current.

14. A method according to claim 13 wherein said current is an AC current.

15. A method according to claim 1, wherein said binding ligand is covalently attached to said solvent accessible transition metal complex.

16. A method according to claim 1, wherein said solvent accessible transition metal complex is covalently attached to said analyte.

17. A method according to claim 1, wherein said ligand is covalently attached to said electrode.

18. A method according to claim 1, wherein said solvent accessible transition metal complex is covalently attached to said electrode.

19. A method according to claim 17, wherein said covalent attachment is via a spacer.

20. A method according to claim 18, wherein said covalent attachment is via a spacer.

21. A method according to claim 19 or 20, wherein said spacer is a conductive oligomer.

22. A method according to claim 1, wherein said analyte is a biomolecule.

23. A method according to claim 22, wherein said biomolecule is selected from the group consisting of proteins, carbohydrates, nucleic acids and lipids.

27. A method according to claim 1 wherein said solvent accessible transition metal complex has at least two coordination sites occupies by a polar coordination group.

29. A method according to claim 1 wherein said conductive oligomer has the formula:

##STR9## wherein Y is an aromatic group;

n is an integer from 1 to 50;

g is either 1 or zero;

e is an integer from zero to 10; and

m is zero or 1;

wherein when g is 1, B--D is a conjugated bond; and

wherein when g is zero, e is 1 and D is preferably carbonyl, or a heteroatom moiety,

wherein the heteroatom is selected from the group consisting of oxygen, sulfur, nitrogen and phosphorus.

30. A method according to claim 29 wherein said aromatic group is phenyl, g is 1, and B--D is an acetylene linkage.

31. A method according to claim 21 wherein said conductive oligomer has the formula:

##STR10## wherein n is an integer from 1 to 50;

m is zero or 1;

C is carbon;

J is carbonyl or a heteroatom moiety, wherein the heteroatom is selected from the group consisting of oxygen, nitrogen, silicon, phosphorus and sulfur; and

G is bond selected from alkane, alkene or acetylene.

32. A method according to claim 22 wherein said biomolecule is a protein.

33. A method according to claim 1 wherein said metal is ruthenium.

34. A method according to claim 1 wherein said metal is osmium.

35. A method according to claim 1 wherein said metal is iron.

36. A method according to claim 1 wherein said metal is platinum.

37. A method according to claim 1 wherein said metal is palladium.

38. A method according to claim 1 wherein said metal is manganese.

39. A method according to claim 1 wherein said metal is technetium.

40. A method according to claim 1 wherein said metal is rhenium.

41. A method according to claim 1 wherein said metal is cobalt.

42. A method according to claim 1 wherein said metal is rhodium.

43. A method according to claim 1 wherein said metal is iridium.

44. A method according to claim 1 wherein said metal is nickel.

45. A method according to claim 1 wherein said metal is copper.

46. A method according to claim 1 wherein said metal is silver.

47. A method according to claim 1 wherein said metal is gold.

24. A method of detecting a target analyte in a test sample comprising:

a) adding an analyte to an electrode comprising;

i) a solvent inhibited transition metal complex comprising a metal selected from the group consisting of manganese, technetium, rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel, palladium, platinum, copper, silver and gold; and

ii) a binding ligand which will bind the target analyte;

such that upon association, a solvent accessible transition metal complex is formed; and

b) detecting a loss of signal as a result of said association.

25. A method according to claim 24 wherein said analyte is an enzyme.

26. A method of detecting a target analyte in a test sample comprising

a) adding an analyte to an electrode comprising:

i) a solvent inhibited transition metal complex comprising a metal selected from the group consisting of manganese, technetium, rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel, palladium, platinum, copper, silver and gold;

ii) a binding ligand which will bind the target analyte; and

iii) an analyte analog;

such that upon association, a solvent accessible transition metal complex is formed; and

b) detecting a loss of signal as a result of said association.

28. A method according to 1 wherein said solvent accessible transition metal complex has at least one coordination site occupied by a water molecule.