MULTI-BIOMARKER BIOSENSOR

• US 20110117582A1
• Filed: 09/02/2010
• Published: 05/19/2011
• Est. Priority Date: 12/01/2005
• Status: Active Grant

• Current Assignee: Northeastern University
• Original Assignee: Northeastern University
• Inventors: TORCHILIN, Vladimir, BUSNAINA, Ahmed, UPPONI, Jaydev, YILMAZ, Cihan, SIAVOSHI, Salome, SOMU, Sivasubramanian, MALIMA, Asanterabi, MUSACCHIO, Tiziana
• Granted Patent: US 8,668,978 B2
• Time in Patent Office: Days
• US Class Current: 435/7.92
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: Y10T 428/2457 Parallel ribs and/or grooves
• CPC Class Codes: Y10T 428/24612 Composite web or sheet
• CPC Class Codes: Y10T 428/24942 including components having same physical characteristic in differing degree

Abstract

Nanosubstrates as biosensors, methods of making such nanosubstrates, and methods of using such nanosubstrates to detect biomarkers are described.

Claims

1. A nanosubstrate comprising:

a substrate layer comprising a photoresist material;

a conductive layer deposited on the substrate layer; and

an insulating layer deposited on the conductive layer, the insulating layer interrupted by one or more nanotrenches or nanowells having a width of at least about 20 nm.

2. The nanosubstrate of claim 1, wherein the nanosubstrate comprises an area of about 0.01 mm² to about 0.1 mm².

3. The nanosubstrate of claim 1, wherein the conductive layer comprises an organic or inorganic conductor.

4. The nanosubstrate of claim 3, wherein the conductive layer comprises gold, aluminum, copper, polyanaline, or a combination thereof.

5. The nanosubstrate of claim 1, wherein the insulating layer comprises a photoresist material.

6. The nanosubstrate of claim 5, wherein the photoresist material is polymethylmethacrylate.

7. The nanosubstrate of claim 1, wherein the thickness of the conductive layer is about 40 nm to about 100 nm.

8. The nanosubstrate of claim 1, wherein the thickness of the insulating layer is about 80 nm to about 150 nm.

9. The nanosubstrate of claim 1, further comprising one or more nanoparticles deposited in the one or more nanotrenches or nanowells.

10. The nanosubstrate of claim 9, wherein the nanoparticles are bound to an antibody or an antigen binding fragment thereof.

11. The nanosubstrate of claim 1, comprising two or more nanotrenches or nanowells having different widths.

12. The nanosubstrate of claim 11, further comprising two or more nanoparticles deposited in said two or more nanotrenches or nanowells, the nanoparticles belonging to two or more different size classes, wherein the nanoparticles in each size class have a diameter less than or equal to the width of the nanotrench or nanowell in which they are deposited.

13. The nanosubstrate of claim 12, wherein the nanoparticles of each size class are covalently bound to a different antibody or antigen binding fragment thereof.

14. A catheter comprising the nanosubstrate of claim 13.

25. A method of detecting a biomarker, comprising

providing the nanosubstrate of claim 13;

and

contacting the nanosubstrate to a biological sample from a subject, the biological sample comprising a biomarker, and wherein binding of the biomarker to the antibody or antigen binding fragment thereof results in a detectable signal, thereby detecting the biomarker.

26. The method of claim 25, wherein the binding of the biomarker to the antibody or antigen binding fragment thereof is determined by ELISA.

27. The method of claim 25, wherein the biomarker is a cancer biomarker.

28. The method of claim 27, wherein detecting the biomarker is indicative of an increased risk of a disease or disorder in the subject.

15. A method of preparing a nanosubstrate, comprising:

providing a first substrate layer;

depositing an adhesion layer onto a surface of the first substrate layer;

depositing a second substrate layer onto the adhesion layer;

depositing a conductive layer onto the second substrate layer;

depositing an insulating layer onto the conducting layer;

creating a pattern of nanotrenches or nanowells in the insulating layer by lithography; and

removing the adhesion layer to release the nanosubstrate, the nanosubstrate comprising the second substrate layer, the conductive layer, and the insulating layer.

16. The method of claim 15, wherein the second substrate layer comprises a photoresist material.

17. The method of claim 15, wherein the steps of depositing the adhesion layer and depositing the conductive layer are carried out by chemical or physical vapor deposition.

18. The method of claim 15, wherein the steps of depositing the second substrate layer and depositing the insulating layer are carried out by spin coating.

19. The method of claim 15, wherein the lithography is electron beam lithography.

20. The method of claim 15, wherein the nanosubstrate comprises an area of about 0.01 mm² to about 0.1 mm².

21. The method of claim 15, further comprising:

contacting the substrate with an aqueous solution of nanoparticles; and

applying a DC voltage between an anode and a cathode, wherein the anode is the conductive layer of the nanosubstrate and the cathode is a conductor placed in the aqueous solution without contacting the nanosubstrate, thereby electrophoretically assembling the nanoparticles within the nanotrenches or nanowells of the nanosubstrate.

22. The method of claim 21, wherein the nanoparticles are covalently bound to an antibody or antigen binding fragment thereof.

23. The method of claim 22, further comprising attaching the nanosubstrate to a catheter.

24. The method of claim 23 wherein the nanosubstrate is attached to the catheter with an adhesive.