Thromb Haemost 1982; 48(03): 294-296
DOI: 10.1055/s-0038-1657284
Original Article
Schattauer GmbH Stuttgart

Purification of Human Tissue-Type Plasminogen Activator in Centigram Quantities from Human Melanoma Cell Culture Fluid and Its Conditioning for Use In Vivo

D Collen
The Center for Thrombosis and Vascular Research, Department of Medical Research, University of Leuven, Belgium
,
D C Rijken
The Center for Thrombosis and Vascular Research, Department of Medical Research, University of Leuven, Belgium
,
J Van Damme
*   Laboratory of Virology, Rega Institute, Department of Human Biology, University of Leuven, Belgium
,
A Billiau
*   Laboratory of Virology, Rega Institute, Department of Human Biology, University of Leuven, Belgium
› Author Affiliations
Further Information

Publication History

Received 18 August 1982

Accepted 13 October 1982

Publication Date:
13 July 2018 (online)

Summary

Human tissue-type plasminogen activator was produced in centigram quantities from 50–60 1 batches of conditioned medium of a human melanoma cell culture. The yields of the procedure remained essentially unchanged during 50 subsequent preparations. The final products were of high purity as assessed by SDS gel electrophoresis.

These materials, after filtration on 0.22 μM Milliporefilters were sterile and free of viruses and pyrogens. Their stability was very good in the fluid form at temperatures up to 56° C. Material obtained by the present procedure has been used to investigate the biological and thrombolytic properties of tissue-type plasminogen activator.

 
  • References

  • 1 Rijken DC, Collen D. Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture. J Biol Chem 1981; 256: 7035-7041
  • 2 Matsuo O, Rijken DC, Collen D. Comparison of the relative fibrinogenolytic, fibrinolytic and thrombolytic properties of tissue plasminogen activator and urokinase in vitro. Thromb Haemostas 1981; 45: 225-229
  • 3 Matsuo O, Rijken DC, Collen D. Thrombolysis by human tissue plasminogen activator and urokinase in rabbits with experimental pulmonary embolus. Nature 1981; 291: 590-591
  • 4 Hoylaerts M, Rijken DC, Lijnen HR, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chem 1982; 257: 2912-2919
  • 5 Rijken DC, Hoylaerts M, Collen D. Fibrinolytic properties of one-chain and two-chain human extrinsic (tissue-type) plasminogen activator. J Biol Chem 1982; 257: 2920-2925
  • 6 Korninger C, Collen D. Neutralization of human extrinsic (tissue-type) plasminogen activator in human plasma: no evidence for a specific inhibitor. Thromb Haemostas 1981; 46: 662-665
  • 7 Korninger C, Stassen JM, Collen D. Turnover of human extrinsic (tissue-type) plasminogen activator in rabbits. Thromb Haemostas 1981; 46: 658-661
  • 8 Korninger C, Collen D. Studies on the specific fibrinolytic effect of human extrinsic (tissue-type) plasminogen activator in human blood and in various animal species in vitro. Thromb Haemostas 1981; 46: 561-565
  • 9 Korninger C, Matsuo O, Suy R, Stassen JM, Collen D. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in dogs with femoral vein thrombosis. J Clin Invest 1982; 69: 573-580
  • 10 Collen D, Stassen JM, Verstraete M. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in rabbits with experimental jugular vein thrombosis. Haemostasis. 1982 11, suppl. 1: Abstract # 57.
  • 11 Weimar W, Stibbe J, Van Seyen AJ, Billiau A, De Somer P, Collen D. Specific lysis of an iliofemoral thrombus by administration of extrinsic (tissue-type) plasminogen activator. Lancet 1981; 2: 1018-1020