Electrode and use thereof

Patent metadata

Field	Value
Publication number	US-10352891-B2
Application number	US-201314412179-A
Country	US
Kind code	B2
Filing date	Jul 2, 2013
Priority date	Jul 3, 2012
Publication date	Jul 16, 2019
Grant date	Jul 16, 2019

Abstract

The application relates to an electrode for use in the electrochemical detection of target species, and in particular for the detection of insulin. The application enables insulin to be detected with high selectivity and with a detection limit in the picomolar range or lower. The electrodes of the invention can easily be reused and are ideally suited for use in point-of-care diagnostics. In a preferred embodiment, the electrode comprises an antibody modified, polyethylene glycol (PEG) monolayer assembled on a gold surface. In a second embodiment, the electrode comprises a chemisorbed zwitterionic carboxybetaine polymer obtained by photopolymerisation of carboxybetaine methacrylate (CBMA). This allows the use of non-Faradaic analysis to determine the insulin concentration even in undiluted blood serum.

First claim

The invention claimed is:

1. A multi-step method of making an electrode for use in an electrochemical impedance spectroscopy technique, the method comprising:
(a) contacting a surface of a substrate with a first composition comprising photopolymerisable monomers to covalently attach the photopolymerisable monomers to the surface of the substrate, thereby obtaining a modified surface having a layer of photopolymerisable monomers covalently attached thereto; and then, after covalent attachment of said layer of photopolymerisable monomers is complete,
(b) contacting said modified surface with a second composition different from the first composition, and comprising further photopolymerisable monomers and optionally crosslinking monomers, and photochemically polymerising the photopolymerisable monomers, thereby generating an electrode comprising polymers disposed on said surface;
wherein the photopolymerisable monomers of step (a) are the same as the further photopolymerisable monomers of step (b).

2. A method according to claim 1 , wherein the photopolymerisable monomers each comprise a photopolymerisable carbon-carbon double bond.

3. A method according to claim 1 , wherein the photopolymerisable monomers are photopolymerisable betaine monomers.

4. A method according to claim 3 , wherein the photopolymerisable betaine monomers each comprise a quaternary ammonium cation and a carboxylate group.

5. A method according to claim 4 , wherein the photopolymerisable betaine monomers have the formula (II)

wherein:
R 1 and R 3 are the same or different and are each a C 1 to C 5 alkylene group;
R 2 and R 2′ are the same or different and are each a C 1 to C 5 alkyl group;
R 4 is a hydrogen atom or a C 1 to C 5 alkyl group; and
X is O or NH.

6. A method according to claim 5 , wherein the photopolymerisable betaine monomers are selected from carboxybetaine methacrylate (CBMA), carboxybetaine acrylamine (CBAA) and carboxybetaine ethylacrylate (CBEA).

7. A method according to claim 1 , which further comprises (c) attaching probe molecules capable of specific binding to a target species to said polymers.

8. A method according to claim 7 , wherein the target species is insulin.

9. A method according to claim 1 , wherein the layer of polymerisable monomers covalently attached to the surface of the substrate in step (a) is a layer of photopolymerisable monomers attached to the surface of a gold substrate via gold-sulfur bonds.

10. A multi-step method of making an electrode for use in an electrochemical impedance spectroscopy technique, the method comprising:
(a) contacting a surface of a substrate with a first composition to covalently attach the first composition to the surface of the substrate, thereby obtaining a modified surface having a layer of the first composition covalently attached thereto;
(b) contacting said modified surface with a second composition, different from the first composition, and comprising photopolymerisable monomers to covalently attach the photopolymerisable monomers to the modified surface, thereby obtaining a further modified surface;
(c) contacting the further modified surface with a third composition comprising additional photopolymerisable monomers and optionally crosslinking monomers, wherein the third composition is different from the second composition; and
(d) photochemically polymerising the photopolymerisable monomers, thereby generating an electrode comprising polymers disposed on said surface;
wherein the photopolymerisable monomers of step (b) are the same as the additional photopolymerisable monomers of step (c).

11. A method according to claim 10 , wherein the photopolymerisable monomers each comprise a photopolymerisable carbon-carbon double bond.

12. A method according to claim 10 , wherein the photopolymerisable monomers are photopolymerisable betaine monomers.

13. A method according to claim 12 , wherein the photopolymerisable betaine monomers each comprise a quaternary ammonium cation and a carboxylate group.

14. A method according to claim 13 , wherein the photopolymerisable betaine monomers have the formula (II)

wherein:
R 1 and R 3 are the same or different and are each a C 1 to C 5 alkylene group;
R 2 and R 2′ are the same or different and are each a C 1 to C 5 alkyl group;
R 4 is a hydrogen atom or a C 1 to C 5 alkyl group; and
X is O or NH.

15. A method according to claim 14 , wherein the photopolymerisable betaine monomers are selected from carboxybetaine methacrylate (CBMA), carboxybetaine acrylamine (CBAA) and carboxybetaine ethylacrylate (CBEA).

16. A method according to claim 10 , further comprising (e) attaching probe molecules capable of specific binding to a target species to the polymers.

17. A method according to claim 16 , wherein the target species is insulin.

18. A method according to claim 10 , wherein the first composition covalently attached to the surface of the substrate in step (a) is attached to a gold substrate surface via gold-sulfur bonds.

Assignees

• Univ Oxford Innovation Ltd

Inventors

• Davis Jason

Classifications

• G01N2333/62

  Insulins · CPC title

• A61B5/1468

  using chemical or electrochemical methods, e.g. by polarographic means · CPC title

• G01N27/026 Primary

  Dielectric impedance spectroscopy (electrochemical impedance spectroscopy for measuring corrosion G01N17/02) · CPC title

• G01N27/3271

  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) · CPC title

• A61B5/1486

  using enzyme electrodes, e.g. with immobilised oxidase · CPC title