Electrochemical biosensor

• US 8,715,981 B2
• Filed: 01/27/2010
• Issued: 05/06/2014
• Est. Priority Date: 01/27/2009
• Status: Active Grant

• Current Assignee: Purdue Research Foundation (Purdue University)
• Original Assignee: Purdue Research Foundation (Purdue University)
• Inventors: Claussen, Jonathan Clay, Franklin, Aaron D., Fisher, Timothy S., Porterfield, D. Marshall
• Primary Examiner(s): Tate, Chris R
• Assistant Examiner(s): Kosar, Aaron J
• Application Number:
  US12/694,876
• Publication Number:
  US 20100285514A1
• Time in Patent Office: 1,560 Days
• Field of Search: None
• US Class Current: 435/174
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C23C 28/322 only coatings of metal elements only
• CPC Class Codes: C23C 28/345 with at least one oxide layer
• CPC Class Codes: G01N 27/3271 Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001

Abstract

Networks of single-walled carbon nanotubes (SWCNTs) decorated with Au-coated Pd (Au/Pd) nanocubes are employed as electrochemical biosensors that exhibit excellent sensitivity (2.6 mA mM⁻¹ cm⁻²) and a low estimated detection limit (2.3 nM) at a signal-to-noise ratio of 3 (S/N=3) in the amperometric sensing of hydrogen peroxide. Biofunctionalization of the Au/Pd nanocube-SWCNT biosensor is demonstrated with the selective immobilization of fluorescently labeled streptavidin on the nanocube surfaces via thiol linking. Similarly, glucose oxidase (GOx) is linked to the surface of the nanocubes for amperometric glucose sensing. The exhibited glucose detection limit of 1.3_M (S/N=3) and linear range spanning from 10 μM to 50 mM substantially surpass other CNT-based biosensors. These results, combined with the structure'"'"'s compatibility with a wide range of biofunctionalization procedures, would make the nanocube-SWCNT biosensor exceptionally useful for glucose detection in diabetic patients and well suited for a wide range of amperometric detection schemes for biomarkers.

Claims

1. An apparatus comprising:

a substrate including an electrically conductive layer and an electrically insulating layer, the insulating layer including a plurality of pores each having a first end and second open end and a passage therebetween, the first end of each pore being open to the conductive layer;

a plurality of carbon nanotubes each having first and second ends, each said nanotube being grown within a different one of said pores, the first end of each said nanotube being in electrical contact with said conductive layer, the second end of each said nanotube extending out of the second open end of the respective pore; and

a plurality of nanoparticles, each said nanoparticle being electrodeposited to the extension of a different one of said nanotubes, each said nanoparticle having bonded to it an enzyme for converting a substance into products

which further comprises a plurality of electrically conductive nanowires, each said nanowire being located within a different one of said pores and providing electrical continuity from the respective nanotube of the pore to said conductive layer.

2. The apparatus of claim 1 wherein said nanoparticle is one of a nanosphere or nanocube.

3. The apparatus of claim 1 wherein said nanoparticle comprises platinum or palladium.

4. The apparatus of claim 1 which further comprises a layer of gold electrodeposited on each said nanoparticle.

5. The apparatus of claim 1 wherein said enzyme is covalently bonded to said nanoparticle.

6. The apparatus of claim 1 wherein said enzyme is bonded to said nanoparticle by a thiol linker.

7. The apparatus of claim 1 wherein the substance is glucose and the enzyme is glucose oxidase.

8. The apparatus of claim 1 wherein the substance is glutamate and the enzyme is glutamate oxidase.

9. The apparatus of claim 1 wherein the substance is glutamate and the enzyme is glutamate dehydrogenase.

10. The apparatus of claim 1 wherein at least one of the products is hydrogen peroxide.

11. The apparatus of claim 1 wherein said insulating layer comprises porous anodic alumina.

12. The apparatus of claim 1 wherein a characteristic overall average length of said nanoparticles is more than about 100 nanometers and less than about 300 nanometers.

13. The apparatus of claim 1 wherein said conductive layer comprises titanium.

14. The apparatus of claim 1 wherein the carbon nanotube is a single walled carbon nanotube.

15. The apparatus of claim 1 wherein the carbon nanotube is a multiwalled walled carbon nanotube.

16. An apparatus comprising:

a substrate including an electrically conductive layer and an electrically insulating layer, the insulating layer including a plurality of pores each having a first end and second open end and a passage therebetween, the first end of each pore being open to the conductive layer;

a plurality of carbon nanotubes each having first and second ends, each said nanotube being grown within a different one of said pores, the first end of each said nanotube being in electrical contact with said conductive layer, the second end of each said nanotube extending out of the second open end of the respective pore; and

a plurality of nanoparticles, each said nanoparticle being electrodeposited to the extension of a different one of said nanotubes, each said nanoparticle having bonded to it an enzyme for converting a substance into products

wherein said substrate includes a catalytic layer placed in between said conductive layer and said insulating layer, and said nanotubes are grown from said catalytic layer.

17. The apparatus of claim 16 wherein said nanoparticle is one of a nanosphere or nanocube.

18. The apparatus of claim 16 wherein said nanoparticle comprises platinum or palladium.

19. The apparatus of claim 16 which further comprises a layer of gold electrodeposited on each said nanoparticle.

20. The apparatus of claim 16 wherein said enzyme is covalently bonded to said nanoparticle.

21. The apparatus of claim 16 wherein said enzyme is bonded to said nanoparticle by a thiol linker.

22. The apparatus of claim 16 wherein the substance is glucose and the enzyme is glucose oxidase.

23. The apparatus of claim 16 wherein the substance is glutamate and the enzyme is glutamate oxidase.

24. The apparatus of claim 16 wherein the substance is glutamate and the enzyme is glutamate dehydrogenase.

25. The apparatus of claim 16 wherein at least one of the products is hydrogen peroxide.

26. The apparatus of claim 16 wherein said insulating layer comprises porous anodic alumina.

27. The apparatus of claim 16 wherein a characteristic overall average length of said nanoparticles is more than about 100 nanometers and less than about 300 nanometers.

28. The apparatus of claim 16 wherein said conductive layer comprises titanium.

29. The apparatus of claim 16 wherein the carbon nanotube is a single walled carbon nanotube.

30. The apparatus of claim 16 wherein the carbon nanotube is a multiwalled walled carbon nanotube.