Enzymatic polymeric sensors

• US 6,673,565 B2
• Filed: 09/14/2001
• Issued: 01/06/2004
• Est. Priority Date: 11/17/1999
• Status: Expired due to Fees

• Current Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: Agentase LLC (Teledyne Technologies Incorporated)
• Inventors: LeJeune, Keith E., Russel, Alan J.
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/952,112
• Publication Number:
  US 20020150959A1
• Time in Patent Office: 844 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
• 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 an analyte enzyme includes at least one substrate compound and at least one indicator compound selected to produce a measurable change of state as a result of the interaction of the substrate and at least one target or analyte enzyme. Each of the indicator(s) and substrate(s) are incorporated within a single polymer.

Claims

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

at least one substrate compound that is selected to react in a reaction catalyzed by the analyte enzyme; and

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

each of the substrate compound 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 as a result of the interaction of the analyte enzyme and the substrate compound.

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

4. The sensor of claim 3 wherein the substrate compound 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 analyte enzyme is a hydrolase enzyme and the dye changes color as a function of pH.

8. The sensor of claim 7 wherein the hydrolase 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 enzyme comprising the steps of:

incorporating at least one substrate compound that is selected to react in a reaction catalyzed by the analyte enzyme; and

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

11. The process of claim 10 wherein the substrate compound 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 substrate compound, the indicator compound and an isocyanate functionalism polyurethane prepolymer.

14. The process of claim 13 wherein the analyte 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 enzyme, the method comprising the steps of:

exposing a sensor to an environment in which the analyte enzyme is present, the sensor including at least one substrate compound that is selected to react in a reaction catalyzed by the analyte enzyme, 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 enzyme and the substrate compound, each of the substrate compound and the indicator compound being incorporated within a single polymer; and

measuring the change of state.