Process And Device For Determining The Activity Of Enzymes In Liquids, Or The Concentration And/Or Activity Of Inhibitors In Liquids

• US 20080268464A1
• Filed: 04/25/2008
• Published: 10/30/2008
• Est. Priority Date: 06/20/1995
• Status: Abandoned Application

• Current Assignee: Papst Licensing GmbH & Company Kg
• Original Assignee: Papst Licensing GmbH & Company Kg
• Inventors: Werle, Bernd, Schumacher, Johannes
• Application Number:
  US12/109,725
• Publication Number:
  US 20080268464A1
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions

Abstract

A process and device are disclosed to determine the activity of enzymes in liquids in a largely automatic manner. The device for carrying out this process has a column (1) with a chromatographic carrier for treating a measurement sample. The carrier is mixed with a substance capable of binding to an enzyme inhibitor present in the measurement sample and that corresponds to at least one enzyme. A measurement sample supply (2) is associated to one end of the column (1). A valve/pump arrangement (7, 11, 14, 15) for filling at least one test tube (5) with a carrier and at least part of the measurement sample is connected downstream of the column (1), in the flow direction of the measurement sample. The carrier is dissociated into cleavage products by the action of the enzyme. The rise in concentration per unit of time of at least one of the cleavage products of the carrier is sensed during an incubation time. As an alternative or supplementary step, the enzyme that corresponds to at least one enzyme inhibitor is extracted by chromatography from a measurement sample to detect enzyme inhibitors in liquids and the thus treated measurement sample is tested for inhibitor concentration and/or activity.

Claims

1-123. -123. (canceled)

124. A system for measuring activity of an enzyme in a liquid sample that includes at least one enzyme and at least one enzyme inhibitor corresponding to said enzyme, the system comprising:

a test vessel containing said sample;

a chromatographic carrier having a substance capable of binding said enzyme inhibitors corresponding to said enzyme in a sample, the substance bound to the carrier effective for binding said enzyme inhibitor with a higher binding affinity than said enzyme, said chromatographic carrier being capable of contacting said sample; and

a means for measuring the activity of said enzyme,

wherein the means for measuring the activity of said enzyme and said test vessel are independently thermostated.

125. The system according to claim 124 wherein the chromatographic carrier contains an excess amount of substance capable of binding said enzyme inhibitor relative to the amount of inhibitor in a sample such that the chromatographic carrier containing the substance capable of binding said enzyme inhibitor can be used repeatedly.

126. The system according to claim 124 wherein the means for measuring activity is a fluorescence detector.

127. The system according to claim 124 wherein the substance bound to the carrier effective for binding an enzyme inhibitor is papain.

128. A method for measuring activity of an enzyme in a liquid sample that includes at least one enzyme and at least one enzyme inhibitor corresponding to said enzyme, the method comprising:

providing said sample to a test vessel;

contacting a said sample with a chromatographic carrier having a substance capable of binding said enzyme inhibitors corresponding to said enzyme in a sample, the substance bound to the carrier effective for binding said enzyme inhibitor with a higher binding affinity than said enzyme;

separating said chromatographic carrier having a substance capable of binding said enzyme inhibitor from said sample; and

measuring the activity of said enzyme with a means for measuring enzyme activity,

wherein the means for measuring the activity of said enzyme and said test vessel are independently thermostated.

129. The method according to claim 128 wherein the chromatographic carrier contains an excess amount of substance capable of binding said enzyme inhibitor relative to the amount of inhibitor in a sample such that the chromatographic carrier containing the substance capable of binding said enzyme inhibitor can be used repeatedly.

130. The method according to claim 128 wherein the means for measuring activity is a fluorescence detector.

131. The method according to claim 128 wherein the substance bound to the carrier effective for binding an enzyme inhibitor is papain.