# US 10,480,091 B2 · Sensors and methods of manufacture thereof

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- Number: US 10,480,091 B2
- Document ID: US10480091B2
- Status: Granted
- Kind: B2
- Application number: 16/373,154
- Filed: 2019-04-02
- Published: 2019-07-25
- Granted: 2019-11-19
- Earliest claimed: 2013-04-16
- Assignees: Purdue Research Foundation
- Applicants: Purdue Research Foundation
- Inventors: Rajtarun Madangopal; Jenna L. Rickus
- Web page: https://patentgrep.com/patent/US10480091B2

## CPC

- C25D 7/00: Electroplating characterised by the article coated
- C09D 5/24: Electrically-conducting paints {(conductive materials H01B1/00)}
- C25D 5/48: After-treatment of electroplated surfaces
- 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)}
- C25D 3/567: {containing more than 50% by weight of platinum group metals}
- G01N 27/3276: {being a hybridisation with immobilised receptors (using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68)}
- C25D: PROCESSES FOR THE ELECTROLYTIC OR ELECTROPHORETIC PRODUCTION OF COATINGS; ELECTROFORMING; APPARATUS THEREFOR
- C09D: COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR (cosmetics A61K; processes for applying liquids or other fluent materials to surfaces, in general, B05D; staining wood B27K5/02; glazes or vitreous enamels C03C; natural resins, French polish, drying-oils, driers, turpentine, per se, C09F; polishing compositions other than French polish, ski waxes C09G; adhesives or use of materials as adhesives C09J; materials for sealing or packing joints or covers C09K3/10; materials for stopping leaks C09K3/12; processes for the electrolytic or electrophoretic production of coatings C25D)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The invention generally relates to sensors, methods of manufacture thereof, methods of use thereof for sensing analytes, such as small molecules and biomolecules, and methods of immobilization. In certain embodiments, the invention provides a multi-analyte sensor. The multi-analyte sensor includes a plurality of sensing electrodes. Each sensing electrode is functionalized with a different molecule (e.g., biomolecule), at least two of the sensing electrodes are spaced apart prior to and after functionalization by 100 μm or less, and there is no cross-talk between the plurality of sensing electrodes.

## Claims

### Claim 1

A method for detecting one or more target biological molecules in a sample, the method comprising:
providing a multi-analyte sensor that comprises a plurality of sensing electrodes, each sensing electrode comprising at least one electrically conductive material and being functionalized with a different enzyme composition from other sensing electrodes of the plurality, wherein at least two of the plurality of sensing electrodes are spaced apart at a microscale level prior to and after functionalization and there is negligible cross-talk between the plurality of sensing electrodes, wherein each sensing electrode comprises: a first layer comprising a first electrically conductive material, a second layer comprising a second electrically conductive material, and a third layer comprising a third electrically conductive material; wherein conductivity of the third electrically conductive material is pH dependent, and wherein the first layer is platinum black, the second electrically conductive biocompatible material is poly(3,4-ethylenedioxythiophene), and the third electrically conductive material is poly(o-aminophenol);
contacting a sample to the multi-analyte sensor; and
detecting one or more target biological molecules from the sample, wherein each of the one or more target biological molecules is detected on a different sensing electrode of the plurality of sensing electrodes.

### Claim 2 (depends on 1)

The method of claim 1, wherein for each sensing electrode respectively, each of the different enzymes is present in the second and third layers.

### Claim 3 (depends on 1)

The method of claim 1, wherein the multi-analyte sensor further comprises a control unit that comprises first and second electrodes, wherein the first electrode is a reference electrode and the second electrode is a counter electrode.

### Claim 4 (depends on 1)

The method of claim 1, wherein a first enzyme is glucose oxidase.

### Claim 5 (depends on 4)

The method of claim 4, wherein a first of the one or more target biological molecules is glucose.

### Claim 6 (depends on 5)

The method of claim 5, wherein a second enzyme is lactate oxidase.

### Claim 7 (depends on 6)

The method of claim 6, wherein a second of the one or more target biological molecules is lactate.

### Claim 8 (depends on 1)

The method of claim 1, wherein the one or more target biological molecules are detected simultaneously.

### Claim 9 (depends on 1)

The method of claim 1, wherein the sensing electrodes align with each other.

### Claim 10 (depends on 1)

The method of claim 1, wherein the sensing electrodes are orthogonal to each other.

### Claim 11

A method for detecting one or more target biological molecules in a sample, the method comprising:
providing a microchip that comprises a plurality of sensing electrodes, each sensing electrode comprising at least one electrically conductive material and being functionalized with a different enzyme composition from other sensing electrodes of the plurality, wherein at least two of the plurality of sensing electrodes are spaced apart at a microscale level prior to and after functionalization and there is negligible cross-talk between the plurality of sensing electrodes, wherein each sensing electrode comprises: a first layer comprising a first electrically conductive material, a second layer comprising a second electrically conductive material, and a third layer comprising a third electrically conductive material; wherein conductivity of the third electrically conductive material is pH dependent, and wherein the first layer is platinum black, the second electrically conductive biocompatible material is poly(3,4-ethylenedioxythiophene), and the third electrically conductive material is poly(o-aminophenol);
contacting a sample to the microchip; and
detecting one or more target biological molecules from the sample, wherein each of the one or more target biological molecules is detected on a different sensing electrode of the plurality of sensing electrodes.

### Claim 12 (depends on 11)

The method of claim 11, wherein for each sensing electrode respectively, each of the different enzymes is present in the second and third layers.

### Claim 13 (depends on 11)

The method of claim 11, wherein the microchip further comprises a control unit that comprises first and second electrodes, wherein the first electrode is a reference electrode and the second electrode is a counter electrode.

### Claim 14 (depends on 11)

The method of claim 11, wherein a first enzyme is glucose oxidase.

### Claim 15 (depends on 14)

The method of claim 14, wherein a first of the one or more target biological molecules is glucose.

### Claim 16 (depends on 15)

The method of claim 15, wherein a second enzyme is lactate oxidase.

### Claim 17 (depends on 16)

The method of claim 16, wherein a second of the one or more target biological molecules is lactate.

### Claim 18 (depends on 11)

The method of claim 11, wherein the one or more target biological molecules are detected simultaneously.

### Claim 19 (depends on 11)

The method of claim 11, wherein the sensing electrodes align with each other.

### Claim 20 (depends on 11)

The method of claim 11, wherein the sensing electrodes are orthogonal to each other.