Method and apparatus for target detection using electrode-bound viruses

• US 9,316,608 B2
• Filed: 07/12/2013
• Issued: 04/19/2016
• Est. Priority Date: 03/09/2006
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Weiss, Gregory A., Penner, Reginald M., Tam, Phillip Y., Yang, Li-Mei, Brigham, Tyler
• Primary Examiner(s): BROWN, MELANIE YU
• Application Number:
  US13/941,231
• Publication Number:
  US 20140014536A1
• Time in Patent Office: 1,012 Days
• US Class Current: 1/1
• CPC Class Codes: G01N 27/327 Biochemical electrodes , e.g. electrical or mechanical details for in vitro measurements
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48

Abstract

A biosensor that is capable of detecting the presence and/or concentration of an analyte or biomarker includes at least one electrically conductive electrode operatively coupled to an impedance analyzer for measuring the change in the resistive impedance ΔZ_Re of the electrode in response to an applied alternating current at a plurality of frequencies. In one embodiment, at least one electrode is covered with a self-assembled monolayer that is chemically bonded to a surface. A plurality of virus particles such as phage viruses are immobilized on the self-assembled monolayer and may be exposed to a test or sample solution. The virus particles may be obtained from phage-displayed libraries to detect a wide variety of targets including, for example, DNA, RNA, small molecules, and proteins or polypeptides. In another embodiment, the virus particles are electrostatically bound to a substrate in between a pair of elongated electrodes disposed on a substrate.

Claims

1. A biosensor comprising:

an electrically conductive gold electrode operatively coupled to an impedance analyzer for measuring the change in the resistive impedance Δ

Z_Re of the electrode;

a self-assembled monolayer chemically bonded to a surface of the electrically conductive gold electrode via a thiol-gold bond formed between N-hydroxysuccinimide thioctic ester and the gold electrode; and

a plurality of phage virus particles covalently bound to the self-assembled monolayer.

2. The biosensor of claim 1, further comprising one or more macromolecules disposed in the self-assembled monolayer.

3. The biosensor of claim 2, wherein the macromolecule comprises bovine serum albumin (BSA) disposed in the self-assembled monolayer.

4. The biosensor of claim 1, wherein the plurality of phage particles comprise bacteriophage M13.

5. The biosensor of claim 1, wherein the plurality of phage particles selectively bind to prostate specific membrane antigen (PSMA).

6. The biosensor of claim 1, further comprising a processor operatively connected to the impedance analyzer for calculating an analyte concentration based at least in part on the measured change in resistive impedance Δ

Z_Re.

7. The biosensor of claim 1, wherein substantially the entire self-assembled monolayer is pinhole free.

8. The biosensor of claim 1, wherein the impedance analyzer is operatively coupled to the electrically conductive gold electrode so as to apply an alternating current within the frequency range of about 2 kHz to about 500 kHz.

9. A system for detecting the presence of an analyte in a sample comprising:

an electrically conductive gold electrode operatively coupled to an impedance analyzer for measuring the change in the resistive impedance Δ

Z_Re of the gold electrode in response to an applied alternating current;

a self-assembled monolayer chemically bonded to a surface of the electrically conductive gold electrode via a thiol-gold bond formed between N-hydroxysuccinimide thioctic ester and the gold electrode;

a plurality of phage virus particles covalently bound to the self-assembled monolayer; and

at least one processor operatively coupled to the impedance analyzer for comparing the change in the measured resistive impedance Δ

Z_Re with one or more pre-set values, the at least one processor configured for outputting a signal indicative of analyte detection based at least in part on the comparison.

10. The system of claim 9, wherein the electrically conductive gold electrode is disposed in a flow cell.

11. The system of claim 9, wherein the at least one processor is configured for outputting a signal indicative of the analyte concentration based at least in part on the comparison.

12. The system of claim 9, wherein the electrically conductive gold electrode comprises an array of electrodes.

13. The system of claim 9, wherein the electrically conductive gold electrode is disposed in a probe.