Hamostaseologie 2005; 25(03): 286-292
DOI: 10.1055/s-0037-1619658
Original Article
Schattauer GmbH

Chromogene Methode zur funktionellen TFPI-Bestimmung

Chromogenic assay for functional determination of TFPI
S. Alban
1   Pharmazeutisches Institut, Christian-Albrechts-Universität zu Kiel
› Author Affiliations
Further Information

Publication History

Publication Date:
27 December 2017 (online)

Zusammenfassung

Aufgrund der vielfältigen TF(tissue factor)-Funktionen ist auch sein natürlicher Gegenspieler, der TFPI (tissue factor pathway inhibitor) von Interesse. Zur quantitativen TFPIBestimmung kann die Antigenkonzentration oder die TFPIAktivität gemessen werden. Methode: Es wird ein chromogener Test zur funktionellen Bestimmung des TFPI vorgestellt und für die Anwendung im Routinelabor modifiziert. Anhand der Testung von über 700 Proben aus einer Heparinstudie mit Gesunden wurde seine Praxistauglichkeit geprüft. Mit der beschriebenen Methode werden sowohl die Aktivität von freiem als auch Lipoprotein-assoziiertem TFPI erfasst. Zusammenfassung: Der chromogene Test erwies sich als einfach und schnell durchführbar und zeichnet sich durch eine hohe Reproduzierbarkeit aus. Insbesondere für die Bestimmung von TFPI in Plasmaproben nach Heparingabe bietet er aufgrund der exzellenten Korrelation eine schnellere und ökonomischere Alternative zur Bestimmung der Gesamt-TFPI-Antigenkonzentration mittels ELISA.

Summary

Due to the manifold functions of tissue factor (TF), also tissue factor pathway inhibitor (TFPI), its endogenous antagonist, is of interest. For the determination of TFPI, either its antigen concentration or its activity can be measured. Method: A chromogenic assay for the functional determination of TFPI is presented and modified for its application in the routine laboratory. Its suitability for daily use was evaluated by testing more than 7000 plasma samples from a heparin study in healthy volunteers. In addition, our data confirm that the method records the activity of both free- and lipoprotein-associated TFPI. Conclusion: The chromogenic assay proved to be rapid and easy to perform and features a high reproducibility. Due to excellent correlation of the results with the total TFPI antigen concentrations, the method represents a more rapid and economic alternative to the determination of total TFPI antigen by ELISA in plasma samples after heparin application.

 
  • Literatur

  • 1 Abildgaard U. Heparin/low molecular weight heparin and tissue factor pathway inhibitor. Haemostasis 1993; 23 (Suppl. 01) 103-6.
  • 2 Alban S. MW-dependent influence of heparin on the form of TFPI circulating in plasma. Semin Thromb Hemost 2001; 27: 503-12.
  • 3 Alban S, Gastpar R. In vitro influence of heparin on plasma total and free tissue factor pathway inhibitor (TFPI) as measured by ELISA. Pharm Pharmacol Lett 2000; 10: 51-4.
  • 4 Alban S, Gastpar R. Plasma levels of total and free tissue factor pathway inhibitor (TFPI) as individual pharmacological parameters of various heparins. Thromb Haemost 2001; 85: 824-9.
  • 5 Alban S, Harenberg J. Neutralization of heparins and heparin-related-oligosaccharides by protamine and polybrene. Pharm Pharmacol Lett 1991; 1: 37-40.
  • 6 Altinbas M, Coskun HS, Er O. et al. A randomized clinical trial of combination chemotherapy with and without low molecular weight heparin in small cell lung cancer. J Thromb Haemost 2004; 2: 1266-71.
  • 7 Bendz B, Anderson TO, Sandset PM. A new sensitive chromogenic substrate assay of tissue factor pathway inhibitor type 1. Thromb Res 2000; 97: 463-72.
  • 8 Bokarewa MI, Morrissey JH, Tarkowski A. Tissue factor as a proinflammatory agent. Arthritis Res 2002; 4: 190-5.
  • 9 Broze Jr. GJ. Tissue factor pathway inhibitor and the revised theory of coagulation. Annu Rev Med 1995; 46: 103-12.
  • 10 Broze Jr GJ, Lange GW, Duffin KL. et al. Heterogeneity of plasma tissue factor pathway inhibitor. Blood Coagul Fibrinol 1994; 5: 551-9.
  • 11 Hansen JB, Huseby KR, Huseby NE. et al. Effect of cholesterol lowering on intravascular pools of TFPI and its anticoagulant potential in type II hyperlipoproteinemia. Arterioscler Thromb Vasc Biol 1995; 15: 879-85.
  • 12 Jeske W, Hoppensteadt D, Fareed J. et al. Measurement of functional and immunological levels of tissue factor pathway inhibitor. Some methodologic considerations. Blood Coag Fibrinol 1995; 6 (Suppl. 01) S73-80.
  • 13 Lindahl AK, Jacobsen PB, Sandset PM, Abildgaard U. tissue factor pathway inhibitor with high anticoagulant activity is increased in post-heparin plasma and in plasma from cancer patients. Blood Coagul Fibrinol 1991; 2: 713-21.
  • 14 Lorenzet R, Maria Benedetta Donati MB. Blood Clotting Activation, Angiogenesis and Tumor Metastasis: Any Role for TFPI?. Thromb Haemost 2002; 87: 928-9.
  • 15 Mousa SA, Mohamed S. Inhibition of endothelial cell tube formation by the low molecular weight heparin, tinzaparin, is mediated by tissue factor pathway inhibitor. Thromb Haemost 2004; 92: 627-33.
  • 16 Novotny WF, Brown SG, Miletich JP. et al. Plasma antigen levels of the lipoprotein-associated coagulation inhbitor in plasma samples. Blood 1991; 78: 387-93.
  • 17 Novotny WF, Palmier M, Wun TC. et al. Purification and properties of heparin-releasable lipoprotein-associated coagulation inhibitor. Blood 1991; 78: 394-400.
  • 18 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res 1988; 50: 803-13.
  • 19 Sandset PM, Larsen ML, Abildgaard U. et al. Chromogenic substrate assay of extrinsic pathway inhibitor (EPI): levels in the normal population and relation to cholesterol. Blood Coag Fibrinol 1991; 2: 425-33.
  • 20 Valentin S, Ostergaard P, Kristensen H. et al. Synergism between full length TFPI and heparin: evidence for TFPI as an important factor for the antithrombotic activity of heparin. Blood Coagul Fibrinolysis 1992; 3: 221-2.