Method and apparatus for measuring temperature changes generated by enzyme activity

• US 4,021,307 A
• Filed: 06/06/1975
• Issued: 05/03/1977
• Est. Priority Date: 06/07/1974
• Status: Expired due to Term

• Current Assignee: Lkb-Produkter AB
• Original Assignee: Lkb-Produkter AB
• Inventors: Mosbach, Klaus H.
• Primary Examiner(s): Tanenholtz, Alvin E.
• Assistant Examiner(s): Fan, C. A.
• Application Number:
  US05/584,595
• Time in Patent Office: 697 Days
• Field of Search: 195/103.5 R, 195/127, 195/115, 195/63, 204/1 E, 204/195 B, 73/15 R
• US Class Current: 435/12
• CPC Class Codes: G01N 25/4873 for a flowing, e.g. gas sample

Abstract

A method and apparatus for measuring temperature changes generated by enzyme activity consists in conducting a matrixbound enzyme through a flowpath formed by a helical coil and sensing the temperature in the immediate vicinity of the flowpath.

Claims

1. Method for measuring enzyme activities comprising the steps of:

a. providing a flow path for a fluid, a portion of said flow path being elongated and having a narrow transverse cross-section compared to the length thereof, said elongated portion being confined within a heat-isolated container;

b. introducing a matrix-bound enzyme preparation into said elongated portion of the flow path;

c. inserting heat-responsive sensor in intimate heat-exchanging relationship to said matrix-bound enzyme in said elongated flow path;

d. producing a flow of fluid containing a substrate which is affected by said enzyme through said flow path, and;

e. determining the enzyme activity solely by temperature changes sensed by said sensor.

2. The method of claim 1, wherein said elongated portion of the flowpath is arranged to surround said sensor.

3. The method defined in claim 2, wherein said elongated portion of the flowpath is arranged to define a helix.

4. The method of claim 2, which includes the step of conducting said fluid containing a substrate through another portion of said flowpath remote from said sensor and in heat-exchanging relationship to a thermostatic path prior to introducing said fluid containing a substrate to said elongated portion of the flowpath.

5. Apparatus for measuring enzyme activity resulting from contact between matrix-bound enzyme particles and a fluid containing a substrate affected by said enzyme, comprising:

a. a closed container immersed in a thermostatic body of liquid;

b. an elongated conduit defining a flow path for said fluid containing a substrate, a first portion of said flow path being in heat-exchanging relationship to said thermostatic body of fluid, a succeeding portion of the flow path being contained within said closed container in heat-isolating relationship thereto;

c. said succeeding portion of the flow path being elongated and having a narrow cross-section with respect to the length thereof;

d. matrix-bound enzyme particles contained in said succeeding portion of the flow path;

e. heat sensor means in heat-exchanging relationship to said matrix-bound enzyme particles, whereby enzyme activity can be determined solely by temperature changes sensed by said sensor when said substrate flows through said flow path.

6. The invention defined in claim 5 wherein a portion of said conduit surrounds said heat sensor to define said succeeding portion of the fluid flow path.

7. The invention defined in claim 6 wherein said portion of the conduit which surrounds the sensor comprises a tightly wound helix defining the side wall of a vessel, one end of the helix being closed, and a heat-exchanging liquid being contained with said heat sensor in said vessel.