Card-based biosensor device

• US 6,300,141 B1
• Filed: 03/02/2000
• Issued: 10/09/2001
• Est. Priority Date: 03/02/1999
• Status: Expired due to Term

• Current Assignee: Helix Biopharma Corp
• Original Assignee: Helix Biopharma Corp
• Inventors: Segal, Donald, Chao, Heman, Wong, Wah Y., McElroy, Jerry
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US09/518,178
• Time in Patent Office: 586 Days
• Field of Search: 204/228.1, 204/229.8, 204/230.2, 204/400, 204/403, 204/406, 204/407, 204/409, 204/422, 349/1, 349/2, 349/33, 435/4, 435/6, 435/7.1, 435/7.5, 435/7-2-, 435/174, 435/176, 435/287.1, 435/287.2, 435/288.7, 436/86, 436/164, 436/518, 436/524, 436/527, 436/536, 422/68.1
• US Class Current: 435/287.1
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

A diagnostic card device for use in detecting or quantitating an analyte present in a liquid sample, comprising a card substrate having a sample introduction region, a biosensor, and a sample-flow pathway communicating between the sample-introduction region and the biosensor, circuitry for generating an analyte-dependent electrical signal from the biosensor; and a signal-responsive element for recording such signal. In one embodiment, the biosensor includes a detection surface with surface-bound molecules of a first charged, coil-forming peptide capable of interacting with a second, oppositely charged coil-forming peptide to form a stable α-helical coiled-coil heterodimer, where the binding of the second peptide to the first peptide, to form such heterodimer, is effective to measurably alter a signal generated by the biosensor. The sample-flow pathway contains diffusibly bound conjugate of the second coil-forming peptide and the analyte (or an analyte analog) and immobilized analyte-binding agent. The analyte in the liquid sample and the conjugate compete for binding with the immobilized analyte-binding agent. Unbound conjugate migrates by capillarity to the biosensor. Liquid sample containing conjugate migrates in the sample flow pathway by capillary action or is driven by a micro-pump. In another embodiment, the biosensor includes an electrode substrate coated with a high-dielectric hydrocarbon-chain monolayer, and having analyte-binding agent attached to the exposed monolayer surface. Binding of analyte to the monolayer-bound analyte-binding agent, and the resultant perturbation of the monolayer structure, causes ion-mediated electron flow across the monolayer.

Claims

1. A diagnostic card device for use in detecting or quantitating an analyte present in a liquid sample, comprising,

a card substrate having formed therein a sample introduction region, a biosensor, and a sample-flow pathway communicating between said sample-introduction region and said biosensor,

circuitry for generating an analyte-dependent electrical signal from the biosensor;

and,

a signal-responsive element for recording said signal,

wherein said biosensor has a detection surface with surface-bound molecules of a first charged, coil-forming peptide for interacting with a second, oppositely charged coil-forming peptide to form a stable α

-helical coiled-coil heterodimer, where the binding of the second peptide to the first peptide, to form said heterodimer, is effective to measurably alter a signal generated by the biosensor, and said sample-flow pathway contains (i) a conjugate of (ia) the second coil-forming peptide and (ib) the analyte or an analyte analog, and (ii) an analyte-binding agent immobilized within the pathway, wherein sample introduced in said sample introduction region is carried through the sample-flow pathway, where the analyte competes with said conjugate for binding to said analyte-binding agent, such that the presence of increasing amounts of analyte leads to increasing amounts of unbound conjugate which is then free to react with said first peptide in said biosensor to alter the signal generated by the biosensor in proportion to the amount of analyte present.

2. The device of claim 1, which further includes a background control biosensor and a control sample-flow pathway connecting the sample-introduction region to the background control biosensor, wherein said control sample-flow pathway does not include said conjugate.

3. The device of claim 1, which further includes a positive control biosensor not connected to the sample-introduction region and which includes a conjugate of (a) the second coil-forming peptide and (b) the analyte or an analyte analog in an amount sufficient to give a maximum biosensor response.

4. The device of claim 1, for use in detecting or quantitating a plurality of different analytes, which further includes, for each analyte, (i) a separate biosensor, and (ii) a separate sample-flow pathway connecting the sample-introduction region to each associated biosensor, where each sample-flow pathway includes (a) a conjugate of the second coil-forming peptide and one of the target analytes or analogs thereof, and (b) an associated analyte-binding agent.

5. The device of claim 4, wherein said sample introduction region comprises a single port communicating with each of the sample-flow pathways.

6. The device of claim 4, wherein said sample introduction region, sample-flow pathways and biosensors are microfabricated on said substrate.

7. The device of claim 1, wherein the sample-flow pathway includes a mixing zone containing the conjugate in releasable form, and a reaction zone containing the analyte-binding agent in immobilized form.

8. The device of claim 1, wherein the biosensor includes a conductive detection surface, a monolayer composed of hydrocarbon chains anchored at their proximal ends to the detection surface, and the first charged coil-forming peptide also anchored to said surface, where the binding of the second peptide to the first peptide, to form said heterodimer, is effective to measurably alter current flow across the monolayer mediated by a redox ion species in an aqueous solution in contact with the monolayer, relative to electron flow observed in the presence of the first peptide alone.

9. The device of claim 8, wherein said circuitry measures current flow across the detection surface, and output a signal related to measured current flow.

10. The device of claim 1, for use in detecting, in a sample, the presence or amount of an analyte, wherein the analyte binds with said analyte-binding agent to form an analyte/analyte-binding agent pair selected from the group consisting of antigen-antibody, hormone-receptor, drug-receptor, cell surface antigen-lectin, biotin-avidin, and complementary nucleic acid strands.

11. The device of claim 1, wherein said signal-responsive element comprises a liquid crystal display device.

12. The device of claim 11, wherein said liquid crystal display device comprises a linear color display device that responds to voltage level from said circuitry.

13. The device of claim 1, wherein said signal-responsive element comprises a magnetic recording medium effective to store a measured signal value from said circuitry.

14. The device of claim 13 for use with a magnetic reader device designed to read the measured signal value stored on said medium.

15. The device of claim 1, wherein said signal-responsive element is separable from portions of the card that are adapted to contact said sample, to provide a sample-free storage record of sample diagnostic results.

16. The device of claim 15 wherein said signal-responsive element comprises a magnetic recording medium effective to store a measured signal value from said circuitry.