Method for determining activated coagulation factors in plasma and plasma derivatives

• US 6,124,110 A
• Filed: 12/13/1999
• Issued: 09/26/2000
• Est. Priority Date: 04/09/1997
• Status: Expired due to Fees

• Current Assignee: DRK-Blutspendedienst Nordrhein-Westfalen GmbH
• Original Assignee: Drk-Blutspendedienst Nordrheinwestfalen Ggmbh
• Inventors: Gesien, Beate, Wober, Guenter
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/402,332
• Time in Patent Office: 288 Days
• Field of Search: 435/13, 435/4
• US Class Current: 435/13
• CPC Class Codes: C12Q 1/56 involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors

Abstract

The invention relates to a method for determining activated coagulation factors in plasma or plasma derivatives. A sample of the plasma or plasma derivative to be examined is incubated with an activated prothrombin complex and procoagulative phospholipid vesicles, the latter containing integrated tissue factor, and thrombin formation is initiated by addition of an appropriate amount of Ca ions. After a determined incubation time, thrombin formation is terminated and the amount of thrombin formed is determined by known methods.

Claims

1. A method for determining activated clotting factors in plasma and plasma derivatives, using the following processing steps:

a) incubating a sample amount of plasma or plasma derivative with activated prothrombin complex and procoagulating phospholipid vesicles, the latter containing integrated tissue factor, and the thrombin generation is initiated by adding a suitable amount of Ca²⁺ ions,

b) after a specified incubation time of 5 min and 10 min, the thrombin generation is ended and the amount of generated thrombin is measured using known methods.

2. The method set forth in claim 1, wherein the content of factor Xa in the activated prothrombin complex is about 1% of the total factor X content with a non-measurable content of factor IIa.

3. The method set forth in claim 1, wherein the tissue factor is produced from biogenic starting material.

4. The method set forth in claim 3, wherein the tissue factor is extracted from bovine brain with a detergent-containing solution of dry acetone powder.

5. The method set forth in claim 1, wherein the tissue factor is integrated in procoagulating phospholipid vesicles.

6. The method set forth in claim 4, wherein the tissue factor is mixed in a detergent-containing solution with phosphatidylserine and phosphatidylcholine and integrated in the procoagulating phospholipid vesicles by dialysis.

7. The method set forth in claim 5, wherein the tissue factor is mixed in a detergent-containing solution with phosphatidylserine and phosphatidylcholine and integrated in the procoagulating phospholipid vesicles by dialysis.