Systems for electrophoretic transport and detection of analytes

• US 6,290,839 B1
• Filed: 08/14/1998
• Issued: 09/18/2001
• Est. Priority Date: 06/23/1998
• Status: Expired due to Fees

• Current Assignee: Osmetech Technology Inc. (Roche Holding AG)
• Original Assignee: Clinical Micro Sensors Inc. (Roche Holding AG)
• Inventors: Kayyem, Jon Faiz, Blackburn, Gary, OʼConnor, Stephen D.
• Primary Examiner(s): Tung, T.
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/134,058
• Time in Patent Office: 1,131 Days
• Field of Search: 204/403, 204/415, 204/409, 204/451, 204/456, 204/601, 204/606, 204/452, 204/602, 204/450, 204/600, 422/68.1, 435/6, 435/183, 205/777.5, 205/775
• 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 2565/107 Alteration in the property of hybridised versus free label oligonucleotides
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides

Abstract

The invention relates to compositions and methods useful in the electrophoretic transport of target analytes to a detection electrode comprising a self-assembled monolayer (SAM). 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) placing said sample in an electric field between at least a first electrode and at least a second electrode sufficient to cause electrophoretic transport of said sample to the vicinity of a detection electrode comprising a self-assembled monolayer, wherein said target analyte directly or indirectly comprises at least one electron transfer moiety (ETM);

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

2. A method according to claim 1 wherein said target analyte directly comprises at least one ETM.

3. A method according to claim 1 wherein said target analyte is directly or indirectly bound to a label probe comprising at least one ETM.

4. A method according to claim 3 wherein at least said first electrode comprises a permeation layer.

5. A method according to claim 1 wherein said detection electrode further comprises a capture binding ligand capable of directly or indirectly binding said target analyte.

6. A method according to claim 1 wherein said detection electrode is the same as said first electrode.

7. A method according to claim 1 wherein said detection electrode is separate from said first or second electrodes.

8. A method according to claim 1 wherein said target analyte is selected from the group consisting of proteins and nucleic acids.

9. A method of detecting a target nucleic acid sequence in a sample comprising:

a) placing said sample in an electric field between at least a first electrode and at least a second electrode sufficient to cause electrophoretic transport of said sample to the vicinity of a detection electrode comprising a self-assembled monolayer, wherein said target sequence directly or indirectly comprises at least one electron transfer moiety (ETM);

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

10. A method according to claim 9 wherein said target sequence directly comprises at least one ETM.

11. A method according to claim 9 wherein said target sequence is directly or indirectly bound to a label probe comprising at least one ETM.

12. A method according to claim 11 wherein said label is positively charged.

13. A method according to claim 11 wherein said label probe comprises a plurality of electron transfer moieties.

14. A method according to claim 11 wherein said label probe is attached to said target sequence by

a) hybridizing a first portion of an amplifier probe to a first portion of said target sequence; and

b) hybridizing at least one amplification sequence of said amplifier probe to said first portion of at least one label probe.

15. A method according to claim 11 wherein said label probe is attached to said target sequence by

a) hybridizing a first portion of a first label extender probe to a first portion of a target sequence;

b) hybridizing a second portion of said first label extender probe to a first portion of an amplifier probe;

c) hybridizing at least one amplification sequence of said amplifier probe to said first portion of at least one label probe.

16. A method according to claim 11 wherein said label probe is attached to said target sequence by

a) hybridizing a first portion of a first label extender probe to a first portion of a target sequence;

b) hybridizing a second portion of said first label extender probe to a first portion of an amplifier probe;

c) hybridizing a first portion of a second label extender probe to a second portion of a target sequence;

d) hybridizing a second portion of said second label extender probe to a first portion of an amplifier probe;

e) hybridizing at least one amplification sequence of said amplifier probe to said first portion of at least one label probe.

17. A method according to claim 9 wherein said detection electrode further comprises a capture probe capable of directly or indirectly binding said target analyte.

18. A method according to claim 17 wherein said target sequence directly hybridizes to said capture probe.

19. A method according to claim 12 wherein a first portion of said target sequence hybridizes to a first capture extender probe, and a second portion of said first capture extender probe hybridizes to a capture probe on the electrode.

20. A method according to claim 17 wherein a first portion of said target sequence hybridizes to a first portion of a first capture extender probe, a second portion of said first capture extender probe hybridizes to a first portion of an capture probe on the electrode, a second portion of said target sequence hybridizes to a first portion of a second capture extender probe, and a second portion of said second capture extender probe hybridizes to a second portion of said capture probe.

21. A method according to claim 17 wherein said label probe is attached to said target sequence by hybridizing said first portion of said label probe to a first portion of said target sequence.

22. A method according to claim 9 wherein said detection electrode is the same as said first electrode.

23. A method according to claim 22 wherein an electroactive charge carrier is used.

24. A method according to claim 9 wherein said detection electrode is separate from said first or second electrodes.

25. A method according to claim 9 wherein at least said first electrode comprises a permeation layer.

26. An electronic device adapted to receive a sample solution comprising a target analyte, said device comprising:

a) a first electrode comprising a permeation layer;

b) a second electrode;

c) a detection electrode comprising;

i) a self-assembled monolayer; and

ii) a capture binding ligand capable of binding said target analyte;

wherein said first and said second electrode are in electrical contact upon the introduction of said sample solution such that electrophoretic transport of said charged target analytes to said detection electrode will occur.

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

a) placing said sample in an electric field between at least a first electrode and at least a second electrode sufficient to cause electrophoretic transport of said sample to the vicinity of a detection electrode comprising a self-assembled monolayer, wherein said target analyte directly comprises at least one electron transfer moiety (ETM);

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

28. A method of detecting a target nucleic acid sequence in a sample comprising:

a) placing said sample in an electric field between at least a first electrode and at least a second electrode sufficient to cause electrophoretic transport of said sample to the vicinity of a detection electrode comprising a self-assembled monolayer, wherein said target sequence directly comprises at least one electron transfer moiety (ETM);

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