Mammalian Cell Expression and Characterization of Recombinant Human Antithrombin III

  • G. Zettlmeissl
  • H. Karges
  • U. Eberhard
Part of the Developments in Hematology and Immunology book series (DIHI, volume 21)


Antithrombin III (AT III) is one of the primary inhibitors of hemostasis [1]. By binding to and inhibiting thrombin, as well as several other activated clotting components (most notably Factors IXa, XIa and XIIa) AT III indirectly influences fibrin clot formation [1]. AT III makes a stochiometric 1:1 complex with thrombin. The complex building rate is increased by the binding of heparin by two orders of magnitude [2]. The physiological importance of AT III in preventing excessive coagulation is demonstrated by studies of individuals whose AT III levels are decreased due to hereditary [3–6] or acquired [7–9] deficiency. Human plasma AT III is a single chain glycoprotein of 432 residues containing three intramolecular disulphide bridges and four N-linked carbohydrate chains [10,11].


Chinese Hamster Ovary Cell Fluorescence Emission Maximum Human Antithrombin High Produce Cell Line Human Liver cDNA Library 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Rosenberg RD. Antithrombin III, biochemistry, function, assay and clinical significance. Ann Univ Sarav Med 1983;3:13–4.Google Scholar
  2. 2.
    Damus PS, Hicks M, Rosenberg RD. Anticoagulant action of heparin. Nature 1973;246:355–7.PubMedCrossRefGoogle Scholar
  3. 3.
    Egeberg D. Inherited antithrombin deficiency causing thrombophilia. Thromb Diath Haemorrh 1965;13:516–41.PubMedGoogle Scholar
  4. 4.
    Sas G, Blasko G, Banhegyi D, Jako J, Palos LA. Abnormal antithrombin III (antithombin III Budapest) as a cause of familiar thrombophilia. Thromb Diath Haemorrh 1974;32:105–15.PubMedGoogle Scholar
  5. 5.
    Sorensen PJ, Dyerberg J, Stoffersen E, Jensen MK. Familiar functional antithrombin III deficiency. Scand J Haematol 1980;24:105–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Koide T, Odani S, Takahashi K, Ono T, Sakuragawa N. Antithrombin III Toyama: Replacement of arginine-47 by cysteine in hereditary abnormal antithrombin III that lacks heparin-binding ability. Proc Natl Acad Sci (USA) 1984;81:289–93.CrossRefGoogle Scholar
  7. 7.
    Abildgaard U, Fagerhol MK, Egeberg D. Comparison of progressive antithrom-bin activity and the concentration of three thrombin inhibitors in plasma. Scand J Clin Invest 1970;26:349–54.PubMedCrossRefGoogle Scholar
  8. 8.
    Thaler E, Balzer E, Kopsa H, Pinggera WF. Acquired antithrombin deficiency in patients with glomerular proteinuria. Haemostasis 1978;7:257–72.PubMedGoogle Scholar
  9. 9.
    Fagerhol MK, Abildgaard U. Immunological studies on human antithrombin III. Scand J Haematol 1970;7:10–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Nordeman B, Nystroem C, Björk I. The size and shape of human and bovine antithrombin III. Eur J Biochem 1977;78:195–203.CrossRefGoogle Scholar
  11. 11.
    Franzen LE, Svenssen S. Structural studies on the carbohydrate portion of human antithrombin III. J Biol Chem 1980;255:5090–3.PubMedGoogle Scholar
  12. 12.
    Bock SC, Wion KL, Vehar GA, Lawn RM. Cloning and expression of the cDNA for human antithrombin III. Nucl Acids Res 1982;10:8113–25.PubMedCrossRefGoogle Scholar
  13. 13.
    Prochownik EV, Markham AF, Orkin SH. Isolation of a cDNA clone for human antithrombin III. J Biol Chem 1983;258:8389–94.PubMedGoogle Scholar
  14. 14.
    Chandra T, Stackhouse R, Kidd VJ, Woo SLC. Isolation and sequence characterization of a cDNA clone of human antithrombin III. Proc Natl Acad Sci (USA) 1983;80:1845–8.CrossRefGoogle Scholar
  15. 15.
    Bröker M, Ragg H, Karges H. Expression of human antithrombin III in Saccha-romyces cerevisiae and Schizosacchoromyces pombe. Biochim Biophys Acta 1987;908: 203–13.PubMedGoogle Scholar
  16. 16.
    Zettmeissl G, Ragg H, Karges H. Expression of biologically active human antithrombin III in Chinese hamster ovary cells. Bio/Technology 1987;5:720–5.CrossRefGoogle Scholar
  17. 17.
    Urlaub G, Chasin LA. Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity. Proc Natl Acad Sci (USA) 1980;77:4216–20.CrossRefGoogle Scholar
  18. 18.
    Kaufman RJ, Wasley LC, Spiliotes et al. Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol Cell Biol 1985;5:1750–9.PubMedGoogle Scholar
  19. 19.
    Kaufman RJ, Wasley LC, Furie BC, Shoemaker CB. Expression, purification and characterization of recombinant-carboxylated Factor IX synthesized in Chinese hamster ovary cells. J Biol Chem 1986;261:9622–8.PubMedGoogle Scholar
  20. 20.
    Mory J, Ben-Barak J, Segev D et al. Effective constitutive production of IFN-7 in Chinese hamster ovary cells. DNA 1986;5:181–93.PubMedCrossRefGoogle Scholar
  21. 21.
    Lee F, Mulligan R, Berg P, Ringold G. Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumor virus chimaeric Plasmids. Nature 1981;294:228–32.PubMedCrossRefGoogle Scholar
  22. 22.
    Ringold G, Dieckmann B, Lee F. Co-expression and amplification of dihydrofolate reductase cDNA and Escherichia coli XGPRT gene in Chinese hamster ovary cells. J Mol Appl Genet 1981;1:165–75.PubMedGoogle Scholar
  23. 23.
    Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–5.PubMedCrossRefGoogle Scholar
  24. 24.
    Ouchterlony O. Progr Allergy 1958;5:1–78.Google Scholar
  25. 25.
    Hensen A, Loeliger EA. Antithrombin III assay. Thromb Diath Haemorrh 1963;9(suppl l):18–29.Google Scholar
  26. 26.
    Kirkwood TBL, Barrowcliffe TW, Thomas DP. An international collaborative study establishing a reference preparation for antithrombin III. Thromb Haemost 1980;43:10–5.PubMedGoogle Scholar
  27. 27.
    Schrader J, Züchner C, Köstering H, Scheler F. Chromogenic substrates for the determination of antithrombin III. Ärztl Lab 1986;32:111-4.Google Scholar
  28. 28.
    Zettlmeissl G, Karges H, Eberhard U. Characterization of recombinant human antithrombin III from mammalian cells. (Unpublished data.)Google Scholar

Copyright information

© Kluwer Academic Publishers, Boston 1988

Authors and Affiliations

  • G. Zettlmeissl
  • H. Karges
  • U. Eberhard

There are no affiliations available

Personalised recommendations