Download PDF
Thromb Haemost 1992; 68(03): 310-314
DOI: 10.1055/s-0038-1656371
Original Article
Schattauer GmbH Stuttgart

Effect of Tissue Factor Pathway Inhibitor (TFPI) in the HEPTEST® Assay and in an Amidolytic Anti Factor Xa Assay for LMW Heparin

Hanne I Kristensen
Biopharmaceuticals Research, Novo Nordisk A/S, Gentofte, Denmark
,
Per B Østergaard
Biopharmaceuticals Research, Novo Nordisk A/S, Gentofte, Denmark
,
Ole Nordfang
Biopharmaceuticals Research, Novo Nordisk A/S, Gentofte, Denmark
,
Ulrich Abildgaard
1   Haematological Research Laboratory, Åker Hospital, Oslo, Norway
,
Anne Karin Lindahl
1   Haematological Research Laboratory, Åker Hospital, Oslo, Norway
› Author Affiliations
Further Information

Publication History

Received 05 February 1992

Accepted after revision 17 April 1992

Publication Date:
04 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Both the HEPTEST® and amidolytic anti factor Xa assays are currently being used for heparin activity detection in plasma from patients receiving standard heparin or low molecular weight heparin (LMWH). In this study we have investigated the influence of recombinant and endogenous Tissue Factor Pathway Inhibitor (TFPI) on these assays. The HEPTEST® determinations were performed on an ACL 300 R Clottimer using the APTT program which resulted in a longer incubation time with factor Xa than recommended by the manufacturer. rTFPI added to plasma prolonged the HEPTEST® clotting time markedly, but had only a little effect in the amidolytic assay. Antibodies against TFPI (anti-TFPI) abolished these effects. The effect of adding rTFPI and Logiparin® was additive. When anti-TFPI IgG was added to samples of normal plasma, a statistically significant shortening of the HEPTEST® clotting time was seen. When anti-TFPI was added to plasma samples from volunteers who had received Logiparin® by subcutaneous or intravenous injection, then the HEPTEST® clotting time was shortened considerably. For some samples the clotting time was halved. These experiments show that the HEPTEST® clotting time is prolonged not only by heparin-antithrombin III, but also by TFPI released by heparin injection.

 

  • References

  • 1 Yin ET. A new single stage measurement of heparin by anti-factor Xa. Semin Thromb Hemostas 1985; 11: 243-244
    PubMedGoogle Scholar
  • 2 Bara L, Combe-Tamzali S, Conrad J, Horellow MH, Samama M. Laboratory monitoring of a low molecular weight heparin (Enoxaparin) with a new clotting test (Heptest). Haemostasis 1987; 17: 127-133
    PubMedGoogle Scholar
  • 3 Bara L, Mardiguian J, Samama M. In vitro effect on heptest of low molecular weight heparin fractions and preparations with various anti-IIa and anti-Xa activities. Thromb Res 1990; 57: 585-592
    CrossrefPubMedGoogle Scholar
  • 4 Halbmayer WM, Feichtinger C, Schwarz T, Gschnait F, Strasser A, Prieshing A, Fisher M. Ein einfacher Test zur Bestimmung der Anti-Xa-Aktivität in Plasmen von Patienten nach subkutaner Gabe von Enoxaparin. Wien Klin Wochenschr 1990; 102: 408-413
    PubMedGoogle Scholar
  • 5 Harenberg J, Giese C, Knödler A, Zimmermann R. Comparative study on a new one stage clotting assay for heparin and its low molecular weight derivatives. Haemostasis 1989; 1: 13-20
    PubMedGoogle Scholar
  • 6 Pedersen PC, Østergaard PB, Hedner U, Bergqvist DMätzsch. Logiparin, after intravenous and subcutaneous administration to healthy volunteers. Thromb Res 1991; 61: 477-487
    CrossrefPubMedGoogle Scholar
  • 7 Bratt G, Törnebohm E, Widlung LLockner. Low molecular weight heparin (Kabi 2165, Fragmin): Pharmacokinetics after intravenous and subcutaneous administration in human volunteers. Thromb Res 1986; 42: 613-620
    CrossrefPubMedGoogle Scholar
  • 8 Bergqvist D, Hedner U, Sjörin E, Holmer E. Anticoagulant effect of two types of low molecular weight heparin administered subcutaneously. Thromb Res 1983; 32: 381-391
    CrossrefPubMedGoogle Scholar
  • 9 Bratt G, Törnehohm E, Cronqvist S, Åberg N, Lockner D. A comparison between low molecular weight heparin (Kabi 2165) and standard heparin in the intravenous treatment of deep venous thrombosis. Thromb Haemostas 1985; 54: 813-817
    PubMedGoogle Scholar
  • 10 Abildgaard U, Norrheim L, Larsen AE, Nesvold A, Sandset PM, Ødegaard OR. Monitoring therapy with LMW heparin: A comparison of three chromogenic substrate assay and the Heptest clotting assay. Hameostasis 1990; 20: 193-203
    PubMedGoogle Scholar
  • 11 Levine MN, Planes A, Hirsh J, Goodyear M, Vochelle N, Gent M. The relationship between anti-factor Xa level and clinical outcome in patients receiving Enoxayparine low molecular weight heparin to prevent deep vein thrombosis after hip replacement. Thromb Haemostas 1989; 62: 940-944
    PubMedGoogle Scholar
  • 12 Broze GJ, Girard TJ, Novotny WF. Regulation of coagulation by a multivalent Kunitz-type inhibitor. Biochemistry 1990; 29: 7539-7546
    CrossrefPubMedGoogle Scholar
  • 13 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic pathway inhibitor. Thromb Res 1988; 50: 803-813
    CrossrefPubMedGoogle Scholar
  • 14 Lindahl AK, Abildgaard U, Stokke G. Release of Extrinsic Pathway Inhibitor after heparin injections: increased response in cancer patients. Thromb Res 1990; 59: 651-656
    CrossrefPubMedGoogle Scholar
  • 15 Lindahl AK, Abildgaard U, Larsen ML, Aamodt L-M, Nordfang O, Beck TC. Extrinsic Pathway Inhibitor (EPI) and the post-heparin anticoagulant effect in tissue thromboplastin induced coagulation. Thromb Res 1991; 65: 14
    PubMedGoogle Scholar
  • 16 Lindahl AK, Abildgaard U, Larsen ML, Staalesen R, Hammer AKG, Sandset PM, Mordfang O, Beck TC. Extrinsic pathway inhibitor released to the blood by heparin is a more powerful inhibitor than is recombinant TFPI. Thromb Res 1991; 62: 607-614
    CrossrefPubMedGoogle Scholar
  • 17 Pedersen AH, Nordfang O, Norris F, Wiberg FC, Christensen PM, Moeller KB, Meidahl-Pedersen J, Beck TC, Norris K, Hedner U, Kisiel W. Recombinant human extrinsic pathway inhibitor. J Biol Chem 1990; 265: 16786-16793
    PubMedGoogle Scholar
  • 18 Sandset PM, Abildgaard U, Pettersen M. A sensitive assay of extrinsic pathway inhibitor in plasma and plasma fractions. Thromb Res 1987; 47: 389-400
    CrossrefPubMedGoogle Scholar
  • 19 Day KC, Hoffman LC, Palmier MO, Kretzmer KK, Huang MD, Pyla EY, Spokas E, Broze GJ, Warren TG, Wun TC. Recombinant lipoprotein-associated coagulation inhibitor inhibits tissue thromboplastin induced intravascular coagulation in the rabbit. Blood 1990; 78: 1538-1545
    PubMedGoogle Scholar
  • 20 Nordfang O, Valentin S, Beck TC, Hedner U. Inhibition of extrinsic pathway inhibitor shortens the coagulation time of normal plasma and of hemophilia plasma. Thromb Haemostas 1991; 66: 464-467
    PubMedGoogle Scholar
  • 21 Wildgoose P, Kisiel W. Activation of human factor VII by factor IXa on human bladder carcinoma cells. Blood 1989; 73: 1888-1895
    PubMedGoogle Scholar
  • 22 Teien AN, Lie M, Abildgaard U. Assay of heparins in plasma using a chromogenic substrate for activated factor X. Thromb Res 1976; 8: 549-553
    PubMedGoogle Scholar
  • 23 Walenga JM, Hoppensteadt D, Fareed J. Laboratory monitoring of the clinical effects of low molecular weight heparins. Thromb Res 1991; Suppl XIV 49-62
    PubMedGoogle Scholar
  • 24 Valentin S, Østergaard P, Kristensen H, Nordfang O. Simultaneous presence of tissue factor pathway inhibitor (TFPI) and low molecular weight heparin has a synergistic effect in different coagulation assays. Blood Coagulation Fibrinolysis 1991; 2: 629-635
    CrossrefPubMedGoogle Scholar
  • 25 Wun TC. Lipoprotein-associated coagulation inhibitor (LAC1) is a cofactor for heparin-synergistic anticoagulant action between LAC1 and sulfated polysaccharides. Blood 1992; 79: 430-438
    PubMedGoogle Scholar
  • 26 Lindahl AK, Abildgaard U, Staalesen R. The anticoagulant effect in heparinized blood and plasma resulting from interactions with extrinsic pathway inhibitor. Thromb Res 1991; 64: 155-168
    CrossrefPubMedGoogle Scholar
  • 27 Nordfang O, Bjørn SE, Valentin S, Nielsen LS, Wildgoose P, Beck TC, Hedner U. The C-terminus of tissue factor pathway inhibitor is essential to its anticoagulant activity. Biochemistry 1991; 30: 10371-10376
    CrossrefPubMedGoogle Scholar