USE AND MAKING OF BIOSENSORS UTILIZING ANTIMICROBIAL PEPTIDES FOR HIGHLY SENSITIVE BIOLOGICAL MONITORING

• US 20120156688A1
• Filed: 06/28/2011
• Published: 06/21/2012
• Est. Priority Date: 06/28/2010
• Status: Active Grant

• Current Assignee: The Trustees of Princeton University (Princeton University)
• Original Assignee: The Trustees of Princeton University (Princeton University)
• Inventors: McAlpine, Michael C., Mannoor, Manu Sebastian
• Granted Patent: US 9,029,168 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• 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: G01N 27/126 comprising organic polymers
• CPC Class Codes: G01N 33/56916 Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia
• CPC Class Codes: Y10T 29/49002 Electrical device making

Abstract

A biosensor and method of making are disclosed. The biosensor is configured to detect a target and may include a peptide immobilized on a sensing component, the sensing component having an anode and a cathode. The immobilized peptide may comprise an antimicrobial peptide binding motif for the target. The sensing component has an electrical conductivity that changes in response to binding of the immobilized peptide to the target. The immobilized peptide may bind one or more targets selected from the list consisting of: bacteria, Gram-negative bacteria, Gram-positive bacteria, pathogens, protozoa, fungi, viruses, and cancerous cells. The biosensor may have a display with a readout that is responsive to changes in electrical conductivity of the sensing component. The display unit may be wirelessly coupled to the sensing component. A resonant circuit with an inductive coil may be electrically coupled to the sensing component. A planar coil antenna may be disposed in proximity to the resonant circuit, the planar coil antenna being configured to provide power to the sensing component.

Claims

1. A biosensor configured to detect a target, the biosensor comprising:

a peptide immobilized on a sensing component, the sensing component having an anode and a cathode, the immobilized peptide comprising an antimicrobial peptide binding motif for the target, the sensing component having an electrical conductivity that changes in response to binding of the immobilized peptide to the target.

2. The biosensor of claim 1, wherein the target is a bacterium.

3. The biosensor of claim 2, wherein the target is a Gram-negative bacterium.

4. The biosensor of claim 3, wherein the target is Escherichia coli.

5. The biosensor of claim 4, wherein the target is Escherichia coli O157:

H7.

6. The biosensor of claim 1, wherein the immobilized peptide binds one or more targets selected from the list consisting of:

bacteria, Gram-negative bacteria, Gram-positive bacteria, pathogens, protozoa, fungi, viruses, and cancerous cells.

7. The biosensor of claim 1, further comprising a display unit electrically coupled to the sensing component forming an electric circuit, the display unit having a readout that is responsive to changes in electrical conductivity of the sensing component.

8. The biosensor of claim 1, further comprising a display unit wirelessly coupled to the sensing component, the display unit having a readout that is responsive to changes in electrical conductivity of the sensing component.

9. The biosensor of claim 1, further comprising a power source electrically coupled to the sensing component.

10. The biosensor of claim 1, further comprising a wireless telemetry device electrically coupled to the sensing component.

11. The biosensor of claim 1, further comprising a resonant circuit with an inductive coil electrically coupled to the sensing component.

12. The biosensor of claim 11, further comprising a planar coil antenna disposed in proximity to the resonant circuit, the planar coil antenna being configured to provide power to the sensing component.

13. The biosensor of claim 1, wherein the sensing component has a longest linear dimension of less than 1 mm.

14. The biosensor of claim 13, wherein the sensing component has a longest linear dimension of less than 1 μ

m.

15. The biosensor of claim 7, wherein the display unit uses impedance spectroscopy to determine the electrical conductivity of the electric circuit and change the readout accordingly.

16. The biosensor of claim 1, wherein the sensing component is an interdigitated microelectrode array.

17. The biosensor of claim 1, wherein the sensing component is a graphene substrate.

18. The biosensor of claim 1, wherein the sensing component is a nanowire substrate.

20. The method of claim 17, further comprising forming an electric circuit including the sensing component and a power source.

21. The method of claim 18, further comprising powering the electric circuit with a planar coil antenna disposed in proximity to the electric circuit.

22. The method of claim 17, further comprising determining a concentration of a target based on the electrical conductivity of the sensing component.

24. The method of claim 21, wherein the antimicrobial peptide is modified prior to being immobilized.

25. The method of claim 21, wherein the biosensor is embedded on a bioresorbable substrate.

19. A method of using an immobilized antimicrobial peptide to detect a target in a sample, the method comprising:

exposing the sample to at least one peptide immobilized on a sensing component, the immobilized peptide having an antimicrobial binding motif, the electrical conductivity of the sensing component changing in response to binding of the immobilized peptide with a target;

measuring the electrical conductivity of the sensing component; and

determining the presence or absence of the target in the sample based on the electrical conductivity of the sensing component.

23. A method of making a biosensor configured to detect a target, the method comprising:

selecting an antimicrobial peptide based on its binding affinity for the target;

immobilizing the antimicrobial peptide on a sensing component, the sensing component having an electrical conductivity that changes based on binding to the target;

electrically coupling an anode and a cathode to the sensing component.

26. The method of claim 23, wherein the bioresorbable substrate is selected from a list consisting of:

poly(ethylene terepthalate), poly(imide), poly(ether sulfone), cellulose, paper, silk and silk fibroin.

27. A biosensor comprising:

an antimicrobial peptide immobilized on a microbe, the microprobe having an electrical conductivity that is dependent on a binding state of the binding molecule;

an anode and a cathode coupled to the microprobe;

a display with a readout electrically coupled to the cathode and anode, forming a circuit, the readout being based on the electrical conductivity of the microprobe.

28. A biosensor comprising:

an interdigitated microelectrode array disposed on a bioresorbable substrate;

an anode and a cathode electrically coupled to the interdigitated microelectrode array;

a resonant circuit with an inductive coil electrically coupled to the cathode and anode; and

a wireless telemetry unit electrically coupled to the resonant circuit, forming an electric circuit.

29. The biosensor of claim 28, further comprising:

a binding molecule disposed on the interdigitated microelectrode array, the interdigitated microelectrode having an electrical conductivity that is dependent on a binding state of the binding molecule.

30. The biosensor of claim 29, wherein binding molecule is selected from the list consisting of:

antimicrobial peptides, antibodies, modified antimicrobial peptides, modified antibodies, chimeric peptides containing antimicrobial peptide binding motifs, chimeric peptides containing antibody binding motifs, DNA fragments, RNA fragments, peptide binding motifs, proteins, small molecules and polymers combinations thereof.

31. The biosensor of claim 28, wherein the bioresorbable substrate is selected from a list consisting of:

poly(ethylene terepthalate), poly(imide), poly(ether sulfone), cellulose, paper, silk, silk fibroin and combinations thereof.

32. The biosensor of claim 28, wherein the bioresorbable substrate is placed in contact with a biologic surface.

33. The biosensor of claim 32, wherein the biologic surface is capable of bioresorption.

34. The biosensor of claim 32, wherein the biologic surface is selected from the list consisting of:

teeth, bone, skin, tissue, hair, nail, cornea, gum, tongue, palate, brain, heart, lung, membrane, leaf, root, bark, fur, feather, chiton and scale.

35. The biosensor of claim 28, further comprising:

a receiver unit having a planar coil antenna electrically coupled to a readout, the planar coil antenna being configured to power the electric circuit, enabling the wireless telemetry unit to send a wireless signal corresponding to an electrical conductivity of the circuit to the receiver unit, the readout being responsive to changes in the electrical conductivity of the circuit.

36. The biosensor of claim 28, wherein the binding molecule is contains a glucose-binding motif.