MULTIPLEXED ELECTROCHEMICAL DETECTION SYSTEM AND METHOD

• US 20080210573A1
• Filed: 03/11/2008
• Published: 09/04/2008
• Est. Priority Date: 12/15/2003
• Status: Active Grant

• Current Assignee: Geneohm Sciences Incorporated (Becton, Dickinson & Co)
• Original Assignee: Geneohm Sciences Incorporated (Becton, Dickinson & Co)
• Inventors: Yang, Xing
• Granted Patent: US 8,420,313 B2
• Time in Patent Office: Days
• US Class Current: 205/777.5
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48

Abstract

Multiplexed analysis of molecular structures of samples. A plurality of sample wells is arranged on a substrate. A plurality of electrodes is fabricated on a first side of the substrate. The electrodes are disposed on the side of the substrate exposed to the sample wells. The electrodes include working electrodes, counter electrodes, and optionally include reference electrodes. At least two of the sample wells includes a plurality of working electrodes. The plurality of electrodes is configured to allow electrochemical analysis of the associated sample wells in a multiplexed fashion. The plurality of electrodes is electrically coupled to an interface to a sample analysis system. The interface to the sample analysis system can include contacts or connections. The sample analysis system controls a signal to the electrodes in a multiplexed fashion and performs the electrochemical analysis.

Claims

1-3. -3. (canceled)

4. An electrochemical detection system comprising:

a multiplexer configured to removably couple to a sample array, the multiplexer configured to selectively apply a first electric potential to a working electrode in each of a plurality of sample locations each comprising a plurality of working electrodes, wherein a single electrical connection between the multiplexer and the sample array couples one of the working electrodes in each of the plurality of sample locations to a first electric potential, the multiplexer further configured to selectively apply a second electric potential to a counter electrode; and

an analyzer coupled to the multiplexer and configured to electrochemically detect a probe hybridization based in part on the first electric potential and the second electric potential.

5. The system of claim 4, wherein the analyzer is configured to detect at least one property of the probe.

6. The system of claim 5, wherein the analyzer is configured to detect the property based on coulometry.

7. The system of claim 5, wherein the analyzer is configured to detect the property based on amperometry.

8. A kit for performing an assay, comprising:

a top plate comprising a plurality of open-bottomed wells;

a substrate having zones adapted to match up to the wells of the top plate such that said zones on the substrate can be matched with bottom openings of wells of the top plate to close said openings;

a plurality of working electrodes positioned on said substrate in each of the zones, the working electrodes each being adapted to have at least one polymer probe attached thereto;

a first electrical connection between a first of the plurality of working electrodes in each of the zones, the first electrical connection being configured for interfacing with an analyzer; and

a second electrical connection between a second of the plurality of working electrodes in each of the zones, the second electrical connection being configured for interfacing with the analyzer.

9. The kit of claim 8, wherein the polymer probes are configured to hybridize with one or more of a nucleic acid, a peptide, a polypeptide, a receptor, a ligand, an antibody, and an enzyme.

10. The kit of claim 8, further comprising:

an interface configured to couple to the analyzer, wherein the interface comprises a portion of the first electrical connection, a portion of the second electrical connection, and an electrical connection to a first counter electrode that is associated with each of the plurality of working electrodes of the first zone.

11. The kit of claim 8, further comprising:

a first counter electrode associated with each of the plurality of working electrodes of a first zone, wherein a current between a particular working electrode of the first zone and the first counter electrode is indicative of one or more characteristics of a particular polymer probe attached to the particular working electrode.

12. The kit of claim 11, further comprising:

a second counter electrode associated with each of the plurality of working electrodes of a second zone, wherein a current between a particular working electrode of the second zone and the second counter electrode is indicative of one or more characteristics of a particular polymer probe attached to the particular working electrode.

13. A method for assembling an assay device, comprising:

attaching polymer probes to electrodes located in zones on the surface of a substrate, the substrate comprising an electrical connection between a single working electrode of each zone such that an electric potential applied to the electrical connection will be applied to the single working electrode of each zone; and

then

attaching the substrate to a top plate having open-bottomed wells to form a multi-well plate in which the top plate defines sides of wells and the substrate defines bottoms of the wells, wherein the electrode-containing zones are located such that the electrodes are accessible to fluid added to the wells.

14. The method of claim 13, wherein each zone is associated with a single respective counter electrode.

15. The method of claim 13, wherein each zone is associated with a single respective reference electrode.

16. A method of analyzing samples for hybridization to captured probes in a sample array, the method comprising:

fabricating an electrode configuration comprising a plurality of electrode zones, each of the electrode zones comprising a plurality of working electrodes and a single counter electrode, wherein each working electrode is electrically coupled to a single working electrode in each of the other zones;

bonding the substrate to a well plate comprising a plurality of wells, wherein each of the wells is associated with one of the electrode zones;

attaching probes to at least some of the working electrodes;

introducing one or more samples to the wells;

applying an electrical potential to at least one working electrode in each of the zones;

determining if respective samples have hybridized to respective of the probes in any of the wells based on at least a current between respective of the at least one working electrodes and respective counter electrodes; and

repeating the steps of applying and determining for each of the working electrodes.

17. The method of claim 16, wherein at least one working electrode in each of the zones comprises a single electrode in each of the zones.

18. The method of claim 16, wherein at least one working electrode in each of the zones comprises all of the electrodes in each of the zones.

19. The method of claim 16, wherein the determining comprises sequentially applying a second electrical potential to counter electrodes in respective electrode zones at the same time as the electrical potential is applied to the at least one working electrode.

20. The method of claim 16, wherein the determining comprises concurrently applying a second electrical potential to counter electrodes in each electrode zones at the same time as the electrical potential is applied to the at least one working electrode.

21. The method of claim 16, wherein each electrode zone comprises a reference electrode.

22. The method of claim 16, wherein at least some of the working electrodes comprise carbon electrodes and the probes comprise polymer probes.

23. The method of claim 16, wherein the attaching probes is performed prior to bonding the substrate to the well plate.