Enzyme-containing polymeric sensors

• US 6,291,200 B1
• Filed: 11/17/1999
• Issued: 09/18/2001
• Est. Priority Date: 11/17/1999
• Status: Expired due to Term

• Current Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: Agentase LLC (Teledyne Technologies Incorporated)
• Inventors: LeJeune, Keith E., Russell, Alan J.
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/441,596
• Time in Patent Office: 671 Days
• Field of Search: 435/20, 435/21, 435/22, 435/24, 435/18, 435/15, 435/16, 435/4, 435/283.1, 435/289.1, 422/50
• US Class Current: 435/20
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/46 involving cholinesterase
• CPC Class Codes: C12Q 1/58 involving urea or urease
• CPC Class Codes: G01N 33/521 Single-layer analytical elements

Abstract

A sensor for detecting the presence of at least one analyte includes at least one enzyme that is selected to either (i) catalyze a reaction of the analyte to chemically convert the analyte to a product compound or (ii) be inhibited by the analyte in the presence of a substrate compound. The sensor also includes at least one indicator compound selected to produce a measurable change of state as a result of the interaction of the analyte and the enzyme. Each of the enzyme and the indicator compound are incorporated within a single polymer.

Claims

1. A sensor for detecting the presence of at least one analyte in an environment, the sensor comprising:

at least one enzyme that is selected (i) to catalyze a reaction of the analyte to chemically convert the analyte to a product compound or (ii) to be inhibited by the analyte in the presence of a substrate compound; and

at least one indicator compound selected to produce a measurable change of state as a result of the interaction of the analyte and the enzyme;

each of the enzyme and the indicator compound being incorporated within a single polymer.

2. The sensor of claim 1 wherein the indicator compound is a dye that changes optical properties in the presence of the product compound.

3. The sensor of claim 2 wherein the polymer matrix comprises a polyurethane.

4. The sensor of claim 3 wherein the enzyme is covalently bonded to the polyurethane.

5. The sensor of claim 4 wherein the indicator compound is entrapped within the polyurethane.

6. The sensor of claim 4 wherein the indicator compound is covalently bonded to the polyurethane.

7. The sensor of claim 3 wherein the enzyme is a hydrolase enzyme and the dye changes color as a function of pH.

8. The sensor of claim 7 wherein the hyrolase enzyme is a lipase, a phosphatase, an amylase, a cellulase, a protease a peptidase, a urease or a deaminase.

9. The sensor of claim 8 wherein the hydrolase enzyme is organophosphorus hydrolase, organophosphorus acid anhydrolase, urease, butyrylcholinesterase or acetylcholinesterase.

10. A process for preparing a sensor to detect the presence of at least one analyte comprising the steps of:

incorporating at least one enzyme that is selected (i) to catalyze a reaction of the analyte to chemically convert the analyte to a product compound or (ii) to be inhibited by the analyte in the presence of a substrate compound into a polymer; and

incorporating at least one indicator compound selected to produce a measurable change of state as a result of the interaction of the analyte and the enzyme into the polymer.

11. The process of claim 10 wherein the enzyme and the indicator compound are incorporated into the polymer during synthesis of the polymer matrix.

12. The process of claim 11 wherein the polymer is a polyurethane.

13. The process of claim 12 wherein the polyurethane is formed by mixing water, the enzyme, the indicator compound and an isocyanate functionalize polyurethane prepolymer.

14. The process of claim 13 wherein the enzyme is a hydrolase enzyme and the indicator compound is a dye that undergoes an observable color change as a function of pH.

15. A method for detecting the presence of at least one analyte, the method comprising the steps of:

exposing a sensor to an environment in which the analyte is present, the sensor including at least one enzyme that is selected (i) to catalyze a reaction of the analyte to chemically convert the analyte to a product compound or (ii) to be inhibited by the analyte in the presence of a substrate compound, the sensor further including at least one indicator compound selected to produce a measurable change of state as a result of the interaction of the analyte and the enzyme, each of the enzyme and the indicator compound being incorporated within a single polymer; and

measuring the change of state.