Binding acceleration techniques for the detection of analytes

• US 7,655,129 B2
• Filed: 04/12/2004
• Issued: 02/02/2010
• Est. Priority Date: 06/23/1998
• Status: Expired due to Fees

• Current Assignee: Clinical Micro Sensors Inc. (Roche Holding AG)
• Original Assignee: Osmetech Technology Inc. (Roche Holding AG)
• Inventors: Meade, Thomas J., Creager, Stephen E., Fraser, Scott, O'Connor, Stephen J., Welch, Thomas W., Terbrueggen, Robert H., Vielmetter, Jost G., Blackburn, Gary, Irvine, Bruce D.
• Primary Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US10/823,503
• Publication Number:
  US 20050003399A1
• Time in Patent Office: 2,122 Days
• Field of Search: 2044031-40315, 205/777.5, 205/778, 205/792
• US Class Current: 205/777.5
• 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: C12Q 2565/629 being a microfluidic device

Abstract

The invention relates to compositions and methods useful in the acceleration of binding of target analytes to capture ligands on surfaces. Detection proceeds through the use of an electron transfer moiety (ETM) that is associated with the target analyte, either directly or indirectly, to allow electronic detection of the ETM.

Claims

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

a) adding said sample to a detection chamber comprising a detection electrode comprising a self assembled monolayer and a capture ligand bound to said self assembled monolayer using biotin and streptavidin;

b) mixing said sample such that said target analyte binds to said capture ligand to form an assay complex, wherein said assay complex further comprises at least one electron transfer moiety (ETM); and

c) detecting the presence of said ETM using said detection electrode.

2. A method according to claim 1 wherein each of said detection electrodes is “

sunken”

or “

recessed”

with respect to the chamber, such that the flow of said sample past each of said detection electrodes causes said mixing.

3. The method of claim 1 or 2 wherein said detecting comprises amperommetry.

4. The method of any one of claims 1 wherein said ETM is a hybridization indicator.

5. The method of claim 4 wherein said ETM comprises a transition metal complex.

6. The method of claim 4 wherein said detecting comprises amperommetry.

7. The method of claim 4 wherein said detecting comprises voltammetry.

8. The method of claim 4 wherein said detecting comprises capacitance measurement.

9. The method of claim 4 wherein said detecting comprises impedence measurement.

10. The method of claim 1 or 2 wherein said detecting comprises capacitance measurement.

11. The method of claim 1 or 2 wherein said detecting comprises impedence measurement.

12. The method of claim 1 or 2 wherein said detecting comprises voltammetry.

13. The method of claim 1 or 2 wherein said detecting comprises amperommetry.

15. A method according to claim 1 or 14 wherein said capture ligand comprises a nucleic acid.

16. The method of claim 15 wherein said target analyte is a nucleic acid and said method further comprises amplifying said nucleic acid analyte with PCR prior to binding to said capture ligand.

17. A method according to claim 1 or 14 wherein said mixing is accomplished by applying an AC/DC pulse.

18. A method according to claim 1 or 14 wherein said mixing is accomplished through the use of mixing particles.

19. A method according to claim 18 wherein said mixing particles comprise microparticulate matter.

20. A method according to claim 1 or 14 wherein said mixing is accomplished through the use of an electrophoretic electrode.

21. The method of claim 1 or 14 wherein said monolayer insulates against one or more of nonspecific binding and nonspecific signaling.

22. The method of claim 1 or 14 wherein said detection electrode is present among an array of detection electrodes.

25. The method of claim 1 or 14, wherein said ETM is a ferrocene.

26. The method of claim 1 or 14, wherein said ETM is a redox protein.

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

a) adding said sample to a detection chamber comprising a detection electrode comprising a self assembled monolayer and a capture ligand bound directly or indirectly to said self assembled monolayer;

b) mixing said sample such that said target analyte binds to said capture ligand to form an assay complex, wherein said assay complex further comprises at least one electron transfer moiety (ETM); and

c) detecting the presence of said ETM using said detection electrode, wherein said detection electrode is sunken or recessed with respect to said chamber such that the flow of said sample past said detection electrode causes said mixing.

23. The method of claim 14 wherein said capture ligand is bound to said self-assembled monolayer using biotin and streptavidin.

24. The method of claim 23 wherein said target analyte is a nucleic acid and said method further comprises amplifying said nucleic acid analyte with PCR prior to binding to said capture ligand.