Biosensor for the detection of a biological target, and method for manufacturing the same

Patent metadata

Field	Value
Publication number	US-10794932-B2
Application number	US-201716328247-A
Country	US
Kind code	B2
Filing date	Aug 29, 2017
Priority date	Aug 29, 2016
Publication date	Oct 6, 2020
Grant date	Oct 6, 2020

Abstract

A substrate for the detection of a biological target by Scanning Electrochemical Microscopy (SECM) is provided. The substrate includes: a conductive surface; a probe area provided on the conductive surface, the probe area comprising receptors responsive to the biological target; and a background area provided on the conductive surface, the background area being disposed around the probe area and comprising electroactive compounds providing imaging contrast between the background area and the probe area.

First claim

The invention claimed is:

1. A substrate for the detection of a biological target by Scanning Electrochemical Microscopy (SECM), comprising:
a conductive surface;
a probe area provided on the conductive surface, the probe area comprising receptors responsive to the biological target; and
a background area provided on the conductive surface, the background area being disposed around the probe area and comprising electroactive compounds providing imaging contrast between the background area and the probe area.

2. The substrate of claim 1 , wherein the conductive surface is a gold surface or a carbon surface.

3. The substrate of claim 1 , wherein the electroactive compounds comprise a metal complex.

4. The substrate of claim 3 , wherein the sandwich metal complex is a metallocene.

5. The substrate of claim 1 , wherein the electroactive compounds are provided on the background area at a surface density between 1 molecule/nm 2 to 10 molecules/nm 2 .

6. The substrate of claim 1 , further comprising a linker layer provided on the conductive surface, wherein:
the linker layer connects the receptors to the conductive surface in the probe area, and
the linker layer connects the electroactive compounds to the conductive surface in the background area.

7. The substrate of claim 6 , wherein the linker layer is made of linkers comprising:
a substrate moiety bound to the conductive surface, and
a ligand attachment moiety bound to the receptors or the electroactive compounds.

8. The substrate of claim 7 , wherein the substrate moiety comprises a functional group selected from the group consisting of a thiol, a sulfide and a disulfide.

9. The substrate of claim 7 , further comprising a blocker compound for blocking unreacted ligand attachment moieties.

10. The substrate of claim 1 , wherein the receptors comprise at least one type of Toll-like receptors (TLRs).

11. The substrate of claim 10 , wherein the receptors comprise heterodimers of TLR receptors.

12. The substrate of claim 1 , wherein the probe area comprises a plurality of spots, each of the spots being at least partially surrounded by the background area.

13. A biosensor for detecting a biological target by SECM, comprising the substrate as defined in claim 1 , and an electrode for applying a potential to the probe area and the background area.

14. The method of claim 1 , wherein the electroactive compounds comprise a metal complex.

15. The method of claim 14 , wherein the sandwich metal complex is a metallocene.

16. A method for manufacturing a substrate for conducting SECM measurements, the method comprising:
providing a conductive surface;
attaching receptors to a first area of the conductive surface to obtain a probe area, the receptors being responsive to a biological target; and
attaching electroactive compounds to a second area of the conductive surface located around the first area to obtain a background area, the electroactive compounds providing imaging contrast between the background area and the probe area.

17. The method of claim 16 , wherein the conductive surface is a gold surface or a carbon surface.

18. The method of claim 16 , wherein the electroactive compounds are attached to the second area at a surface density between 1 molecule/nm 2 to 10 molecules/nm 2 .

19. The method of claim 16 , further comprising binding linkers to the conductive surface to obtain a linker layer, wherein:
attaching the receptors to the first area of the conductive surface comprises binding the receptors to the linker layer; and
attaching the electroactive compounds to the second area of the conductive surface comprises binding the electroactive compounds to the linker layer.

20. The method of claim 19 , wherein binding the linkers to the conductive surface comprises:
binding a substrate moiety of the linkers to the conductive surface; and
binding a ligand attachment moiety of the linkers to the receptors in the first area, and to the electroactive compounds in the second area.

21. The method of claim 20 , wherein the substrate moiety comprises a sulfur-bearing functional group selected from the group consisting of a thiol, a sulfide and a disulfide.

22. The method of claim 20 , further comprising binding a blocker compound to unreacted ligand attachment moieties to block the unreacted ligand attachment moieties.

23. The method of claim 16 , wherein the receptors comprise at least one type of Toll-like receptors (TLRs).

24. The method of claim 16 , wherein the probe area comprises a plurality of spots, each of the spots being at least partially surrounded by the background area.

Assignees

• Governing Council Univ Toronto
• Her Majesty The Queen In Right Of Canada As Represented By The Minster Of Nat Defense

Inventors

• She Zhe
• Topping Kristin
• Kraatz Heinz-Bernhard

Classifications

• G01Q60/60 Primary

  SECM [Scanning Electro-Chemical Microscopy] or apparatus therefor, e.g. SECM probes · CPC title

• G01Q30/20 Primary

  Sample handling devices or methods · CPC title