Bridged element for detection of a target substance

• US 9,857,366 B2
• Filed: 10/22/2013
• Issued: 01/02/2018
• Est. Priority Date: 08/06/2003
• Status: Expired due to Fees

• Current Assignee: Bridger Technologies, Inc. (Bio-Rad Laboratories Incorporated)
• Original Assignee: Bridger Technologies, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Albert, Fred, Wright, Brad
• Primary Examiner(s): Lin, Jerry
• Application Number:
  US14/059,826
• Publication Number:
  US 20140141525A1
• Time in Patent Office: 1,533 Days
• Field of Search: 702 27
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2523/307 Denaturation or renaturation by electric current/voltage
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Physical changes resulting from an association between a template molecule and a target molecule are detected by monitoring changes in the template molecule. Exemplary changes include a change in a physical dimension or stiffness of the template molecule, a change in electrical conductivity of the template molecule and a change in the energy required to dissociate the target molecule and the template molecule. The magnitude of the change is indicative of the specific identity of the target molecule.

Claims

1. A device, comprising:

at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the first surface is independent of the second surface, the first contact point and the second contact point having a first physical parameter;

a template molecule linked between the at least two contact points, wherein the template molecule is a selected polypeptide molecule which includes at least one recognition site for a target molecule; and

a detector coupled to the first contact point and the second contact point, the detector configured to generate an output signal based on a change in the first physical parameter indicative of electrical conduction through the template molecule, the detector further comprising a driving circuit,

wherein the template molecule, the first contact point, the detector and the second contact point form an electrical circuit, and further, wherein the driving circuit is configured to provide a current through or voltage across the electrical circuit to disassociate the template molecule from the target molecule.

2. The device of claim 1 wherein the polypeptide is bonded at its carboxy terminus to the first contact point and at its amino terminus to the second contact point.

3. The device of claim 1 wherein the polypeptide is bonded at a point other than a terminus to the first contact point and another point other than a terminus to the second contact point.

4. The device of claim 1 wherein the polypeptide comprises a recognition region for the target molecule and the target molecule is an antibody.

5. The device of claim 1 wherein the polypeptide comprises a recognition site derived from an antibody that associates with the target molecule, the target molecule further comprising an antigenic site for said antibody.

6. The device of claim 1 wherein the polypeptide comprises the biochemical and physical features to permit a molecular association with the target molecule.

7. The device of claim 6 wherein the target molecule is an antagonist or agonist for the polypeptide.

8. The device of claim 1 wherein the target molecule is a second polypeptide that forms a molecular association with at least a portion of the template molecule.

9. The device of claim 1 wherein the target molecule comprises metal ions.

10. The device of claim 1 wherein the first physical parameter includes at least one of:

a) a resonant frequency of the first contact point relative to a reference point;

b) a resonant amplitude of the first contact point relative to the reference point;

c) a distance between the first contact point and the reference point; and

d) an alignment between the first contact point and the reference point.

11. The device of claim 10 wherein the reference point includes the second contact point.

12. The device of claim 1 wherein the first surface includes a movable surface.

13. The device of claim 1 wherein the detector includes at least one of an optical sensor, a magnetic field sensor, an electric field sensor, a capacitance sensor, a resistance sensor and a strain sensor.

14. The device of claim 1 wherein the detector includes at least one of a comparator and a bridge circuit.

15. The device of claim 1 wherein the detector includes a resonance driver in communication with the first surface.

16. The device of claim 1 wherein the first contact point and the second contact point have a second physical parameter, and wherein the detector is configured to generate the output signal based on a change in the second physical parameter, wherein the change in the second physical parameter corresponds to the association of the target molecule and the template molecule.

17. The device of claim 1 wherein the first physical parameter is an electrical parameter.

18. The device of claim 17 wherein the detector includes a voltage source coupled to the first contact point and the second contact point and wherein the first electrical parameter includes a measure of a current.

19. The device of claim 18 wherein the voltage source is configured to supply an increasing potential.

20. The device of claim 17 wherein the detector includes a current source coupled to the first contact point and the second contact point and wherein the first electrical parameter includes a measure of a voltage.

21. The device of claim 20 wherein the current source is configured to supply an increasing current.

22. The device of claim 17 wherein the change in the first physical parameter corresponds to a disassociation of the target molecule and the template molecule induced by the electrical signal delivered by the driver circuit.

23. The device of claim 1 wherein at least one of the first surface and the second surface includes at least one of glass, quartz, silicon, gold, nickel, platinum and a polymer.

24. A method comprising:

exposing a target molecule to a template polypeptide molecule, the template molecule linked between at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the template molecule includes at least one recognition site for the target molecule;

generating an output signal as a function of a change in a first physical parameter indicative of electrical conduction through the template molecule measured using the first contact point relative to a reference point, wherein the change in the first physical parameter corresponds to an association of the target molecule and the template molecule; and

generating a current through or voltage across the first contact point, the template molecule and the second contact point to disassociate the target molecule from the template molecule.

25. The method of claim 24 further including:

monitoring a resonant frequency of the first contact point relative to a reference point;

monitoring a resonant amplitude of the first contact point relative to the reference point;

monitoring a distance between the first contact point and the reference point; and

monitoring an alignment between the first contact point and the reference point.

26. The method of claim 25, wherein the target molecule and the template molecule form a protein-protein or protein conjugate when the target molecule is exposed to the template molecule.

27. The method of claim 25 wherein the reference point includes the second contact point.

28. The method of claim 24 further including applying an electrical signal to the template molecule to disassociate the target molecule, and wherein the change in the first physical parameter corresponds to a disassociation of the target molecule and the template molecule caused by the current or voltage.

29. A system comprising:

a target molecule introduction port to receive a sample;

a sensor having a template molecule in communication with the target molecule introduction port, the template molecule linked between a first point on a first surface and a second point on a second surface, the first surface independent of the second surface and wherein the template molecule is a polypeptide and has at least one recognition site specific to a target molecule;

a detector coupled to the first point and the second contact point and configured to generate an output signal based on a change in a measured parameter of the first point and corresponding to an association of the template molecule with the target molecule, wherein the measured parameter is based on electrical conduction through the template molecule; and

an output circuit to provide a result based on the output signal, wherein the template molecule, the detector and the second contact point form an electrical circuit, and further wherein the detector further comprises a driving circuit configure to provide a current through or voltage across the electrical circuit to disassociate the template molecule from the target molecule.

30. The system of claim 29 further including a plurality of sensors, each sensor coupled to the detector by a multiplexer.

31. The system of claim 29 a processor coupled to the detector and having access to a memory, wherein the memory provides data storage for identifying a target molecule based on the change in the measured parameter.

32. The system of claim 29 wherein the output circuit includes at least one of an interface, a display and a wireless transceiver.

33. The system of claim 29 further including a test circuit coupled to the template molecule to determine conductivity of the template molecule.

34. The system of claim 29 wherein the change in the measured parameter corresponds to a disassociation of the target molecule and the template molecule induced by the electrical signal delivered by the driving circuit.

35. The system of claim 29 further including a housing containing at least one of the target molecule introduction port, the sensor, the detector and the output circuit.