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US 7,923,217 B2Granted
Specificity in the determination of antithrombin
AssigneeRoche Diagnostics Operations, Inc.
InventorAdena Enno
Life of this patent
Estimating the term…
Abstract
Methods and reagents for determining antithrombin III (AT) in body fluids by adding an AT binding partner to the sample and determining the free AT binding partner.
Claims10
A method for determining the antithrombin III (AT) content in a sample that may contain one or more pharmaceutical compounds that inhibit thrombin, the method comprising:
- (a) providing a reaction mixture by contacting the sample with a first reagent R1 comprising thrombin, under conditions wherein the thrombin essentially does not interact with AT but interacts with the one or more pharmaceutical compounds that inhibit thrombin, if present,
- (b) adding to the reaction mixture a second reagent R2 comprising a chromogenic substrate which is a peptide substrate for thrombin, and determining the amount of thrombin activity in the reaction mixture,
- (c) changing the conditions of the reaction mixture by adding to the reaction mixture a third reagent R3 comprising heparin such that thrombin interacts with AT,
- (d) conducting a second determination of the thrombin activity in the reaction mixture, wherein the determination comprises using the reagent R2,
- (e) determining the difference between the first and second determinations of thrombin activity, and
- (f) determining the AT content of said sample from said difference determined in step (e).
3 more elements
A method for determining the amount of antithrombin III (AT) in a sample containing one or more pharmaceutical compounds that inhibit thrombin, the method comprising:
- (a) providing a reaction mixture by mixing the sample with thrombin and an antagonist for heparin under conditions wherein the thrombin essentially does not interact with AT but interacts with the one or more pharmaceutical compounds that inhibit thrombin,
- (b) adding to the reaction mixture a chromogenic substrate which is a peptide substrate for thrombin, and conducing a first determination of the thrombin activity in the reaction mixture,
- (c) changing the conditions of the reaction mixture by adding heparin to the reaction mixture such that thrombin interacts with AT,
- (d) conducting a second determination of the thrombin activity in the reaction mixture using the chromogenic substrate,
- (e) determining the difference between the first and second determinations of thrombin, and
- (f) determining the AT content of said sample from said difference determined in step (e).
3 more elements
Dependencies are inferred from the claim text.