Enzyme-based device for environmental monitoring

• US 20060073490A1
• Filed: 10/06/2004
• Published: 04/06/2006
• Est. Priority Date: 10/06/2004
• Status: Active Grant

• Current Assignee: Flir Detection, Inc. (Teledyne Technologies Incorporated)
• Original Assignee: Agentase LLC (Teledyne Technologies Incorporated)
• Inventors: LeJeune, Keith E., Erbeldinger, Markus, Holzapfel, Paul L., Mysliwczyk, Richard J.
• Granted Patent: US 7,422,892 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/46 involving cholinesterase

Abstract

A sensor for the intermittent or continuous detection of the presence of at least one analyte in an environmental sample 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 sensor for monitoring or 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. Optionally, each of the enzyme and the indicator compound are incorporated within a single polymer.

Claims

1. An enzyme-based environmental monitoring device for detecting a target analyte comprising:

an immobilized enzyme;

a mechanism for delivering an environmental sample to said immobilized enzyme, wherein said environmental sample optionally contains a target analyte;

a mechanism for delivering an enzyme substrate compound and optionally a pH sensitive dye to said immobilized enzyme; and

one or more sensors for detecting the level of enzyme activity of said immobilized enzyme and presence, if any, of said target analyte, wherein said target analyte acts as an enzyme inhibitor in the presence of said immobilized enzyme and said enzyme substrate compound.

2. The device of claim 1 wherein said immobilized enzyme is trapped in an immobilization matrix.

3. The device of claim 2 wherein said immobilized enzyme is covalently bonded to a polymer matrix.

4. The device of claim 1 wherein said immobilized enzyme is a hydrolase enzyme.

5. The device of claim 3 wherein said immobilized enzyme is butyrylcholinesterase and further wherein said target analyte is a cholinesterase inhibitor.

6. The device of claim 2 one wherein said enzyme substrate compound is selected from one or more of a group comprising acetylcholine and butyrylcholine and further wherein said immobilized enzyme is selected from a group comprising is acetylcholinesterase and butyrylcholinesterase.

7. The device of claim 2 wherein said enzyme substrate compound is benzoyl-arginine-ethyl-ester and further wherein said immobilized enzyme is papain.

8. The device of claim 2 wherein said enzyme substrate compound is urea and further wherein said immobilized enzyme is urea aminohydrolase.

9. The device of claim 4 wherein said one or more sensors are selected from a group consisting of one or more pH electrodes and one or more color readers.

10. The device of claim 1 wherein at least one of said environmental sample or said enzyme substrate compound is delivered to said immobilized enzyme intermittently or continuously.

11. The device of claim 1 wherein said environmental sample and said enzyme substrate compound are delivered to said immobilized enzyme simultaneously.

12. The device of claim 1 that is capable of alternating the flow of the delivery of said environmental sample to said immobilized enzyme, and then delivering said enzyme substrate compound to said immobilized enzyme in a repetitive pattern such that either said environmental sample is delivered to said immobilized enzyme for one time period or said enzyme substrate compound is delivered to said immobilized enzyme for another time period.

13. The device of claim 9 wherein said color reader is a RGB color to frequency converter.

14. The device of claim 9 wherein data read from said sensors are recorded for later analysis.

15. The device of claim 14 wherein said data is stored on-board said device.

16. The device of claim 14 wherein said data is directly output from said device.

17. The device of claim 14 wherein said data is wirelessly transmitted by said device.

18. The device of claim 1 wherein said mechanism for delivering said environmental sample is selected from a group comprising pumps and fans.

19. The device of claim 9 further comprising a microcontroller for collecting and analyzing said data and for determining if said target analyte is present in said environmental sample by comparing said data to a known baseline.

20. The device of claim 17 further comprising an alarm for providing notification of the presence of said target analyte in said environmental sample.

21. The device of claim 17 wherein a plurality of sensors are used and further wherein said alarm is activated only if all of said plurality of sensors return consistent data.

23. The device of claim 20 further comprising a color reader for detecting changes in the color of said pH sensitive dye compound at said immobilization matrix.

24. The device of claim 21 further comprising a pH electrode for detecting changes in the pH of said stream of water or air after its passage through said immobilization matrix.

22. An enzyme-based environmental monitoring device for detecting contamination in a sample of water or air comprising:

an immobilized enzyme covalently bonded with a polymer to form an immobilization matrix;

a first pump or fan for continuously delivering a stream of environmental water or air to said immobilization matrix;

a second pump for delivering an enzyme substrate compound and optionally a pH sensitive dye compound to said stream of water or air prior to its delivery to said immobilization matrix; and

one or more sensors for detecting the level of enzyme activity at said immobilization matrix.

25. The device of claim 22 further comprising a computer for reading data output from said sensors and for analyzing said data to determine if said sample of water or air is contaminated.

26. An enzyme-based environmental monitoring device for detecting a target analyte comprising:

a first reaction system including a first enzyme and a substrate for the first enzyme, a mechanism for delivering an environmental sample to the device either intermittently or continuously, wherein said environmental sample optionally contains the analyte, wherein the analyte inhibits the first enzyme; and

at least a second reaction system that reacts to produce a first detectable state only when the first enzyme is inhibited.

27. The enzyme-based environmental monitoring device of claim 26 wherein the reaction of the first reaction system produces a second detectable state different from the first detectable state.

28. The enzyme-based environmental monitoring device of claim 26 wherein the reaction of the first reaction system causes pH to change in a first direction and wherein the reaction of the second reaction system causes pH to change in a second direction, opposite of the first direction.

29. An enzyme-based environmental monitoring device for detecting a target analyte comprising:

at least one enzyme that is selected to catalyze a reaction of the analyte to chemically convert the analyte to a product compound, a mechanism for delivering an environmental sample to the device either intermittently or continuously, wherein the environmental sample optionally contains said analyte, and

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