Download PDF
Thromb Haemost 1979; 42(03): 924-928
DOI: 10.1055/s-0038-1666941
Original Article
Schattauer GmbH Stuttgart

Purification of Urokinase by a Beta-Naphthamidine Affinity Column

B Åstedt
The Research Laboratories of the Departments of Gynecology & Obstetrics and Pediatrics, University of Lund, Malmö Allmänna Sjukhus, 21401 Malmö, Sweden, the Section of Biosciences, Karl-Marx-University of Leipzig, GDR, and the Research Laboratories of Lowens Pharmaceuticals, Denmark
,
L Holmberg
The Research Laboratories of the Departments of Gynecology & Obstetrics and Pediatrics, University of Lund, Malmö Allmänna Sjukhus, 21401 Malmö, Sweden, the Section of Biosciences, Karl-Marx-University of Leipzig, GDR, and the Research Laboratories of Lowens Pharmaceuticals, Denmark
,
G Wagner
The Research Laboratories of the Departments of Gynecology & Obstetrics and Pediatrics, University of Lund, Malmö Allmänna Sjukhus, 21401 Malmö, Sweden, the Section of Biosciences, Karl-Marx-University of Leipzig, GDR, and the Research Laboratories of Lowens Pharmaceuticals, Denmark
,
P Richter
The Research Laboratories of the Departments of Gynecology & Obstetrics and Pediatrics, University of Lund, Malmö Allmänna Sjukhus, 21401 Malmö, Sweden, the Section of Biosciences, Karl-Marx-University of Leipzig, GDR, and the Research Laboratories of Lowens Pharmaceuticals, Denmark
,
J Ploug
The Research Laboratories of the Departments of Gynecology & Obstetrics and Pediatrics, University of Lund, Malmö Allmänna Sjukhus, 21401 Malmö, Sweden, the Section of Biosciences, Karl-Marx-University of Leipzig, GDR, and the Research Laboratories of Lowens Pharmaceuticals, Denmark
› Author Affiliations
Further Information

Publication History

Received 12 September 1978

Accepted 21 November 1978

Publication Date:
23 August 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Urokinase was purified by affinity chromatography using 6-amino-naphthamidine-(2), a new specific ligand based on the urokinase inhibitor β-naphthamidine. Urokinase was firmly bound at pH 7.0 and could be eluted at pH 3.0. The protein which passed the column at pH 7.0 without being bound did not contain any urokinase activity. This is an important property because it can be utilized for raising a monospecific urokinase antiserum by absorbing unspecific antibodies with only a minor loss of antiserum titre.

 

  • References

  • 1 Cuatrecasas P, Anfinsen CB. 1971. Jakoby WB. (ed.) In Methods in Enzyntology, Vol. 22. pp 345-378 Academic Press; New York:
  • 2 Geratz JD, Cheng MC-F. 1975; The inhibition of urokinase by aromatic diamidines. Thrombosis et Diathesis Haemorrhagica (Stuttg.) 33: 230
    PubMedSearch in Google Scholar
  • 3 Hoare DC, Koshland Jr DE. 1967; A method for the quantitative modification and estimation of Carboxylic acid groups in proteins. Journal of Biological Chemistry 242: 2447
    PubMedSearch in Google Scholar
  • 4 Holmberg L, Bladh B, Åstedt B. 1976; Purification of urokinase by affinity chromatography. Biochimica Biophysica Acta 445: 215
    CrossrefPubMedSearch in Google Scholar
  • 5 Landmann H. 1973; Studies on the mechanism of action of synthetic antifibrinolytics. A comparison with the action of derivatives of benzamidine on the fibrinolytic process Thrombosis et Diathesis Haemorrhagica, (Stuttg.) 29: 253
    PubMedSearch in Google Scholar
  • 6 Lesuk A, Terminiello L, Traver JH. 1965; Crystalline human urokinase: some properties. Science 147: 880
    CrossrefPubMedSearch in Google Scholar
  • 7 Maciag T, Weibel MK, Kendall Pye E. 1974. In Jakoby WB, Wilchek M. Eds Methods in Enzymology, Vol. 34. pp 451-459 Academic Press; New York:
  • 8 Nilsson IM, Olow B. 1962; Fibrinolysis induced by streptokinase in man. Acta chirurgica scandinavica 123: 247
    PubMedSearch in Google Scholar
  • 9 Ogawa N, Yamamoto H, Katamine T, Tajima H. 1975; Purification and some properties of urokinase. Thrombosis et Diathesis Haemorrhagica (Stuttg.) 34: 194
    PubMedSearch in Google Scholar
  • 10 Ong EB, Soberano ME, Johnson AJ, Schoellmann G. 1977; Studies on the Biochemistry of urokinase. Thrombosis and Haemostasis (Stuttg.) 38: 801
    Thieme ConnectPubMedSearch in Google Scholar
  • 11 Prentice CR M, Rogers KM, Mcnicol GP. 1973; Evaluation of a new preparation of urokinase. Thrombosis et Diathesis Haemorrhagica 30: 114
    PubMedSearch in Google Scholar
  • 12 Tamura Y, Fujh S. 1976; Purification of human plasma Kallikrein and Urokinase by affinity chromatography. Journal of Biochemistry 80: 507
    PubMedSearch in Google Scholar
  • 13 Wagner G, Richter P, Lischke I. 1978 Pharmazie 33 in press
    PubMed
  • 14 Walton PL. 1967; The hydrolysis of α-N-Acetylglycyl-L-Lysine Methyl Ester by urokinase. Biochimica Biophysica Acta 132: 104
    CrossrefPubMedSearch in Google Scholar
  • 15 White WF, Barlow GH, Mozen MM. 1966; The isolation and characterization of plasminogen activators (urokinase) from human urine. Biochemistry 5: 2160
    CrossrefPubMedSearch in Google Scholar