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Thromb Haemost 1986; 55(03): 347-351
DOI: 10.1055/s-0038-1661561
Original Article
Schattauer GmbH Stuttgart

Monoclonal Antibodies Against Human High Molecular Weight Urinary Urokinase: Application for Affinity Purification of Urinary Prourokinase[*]

J Wojta
The Laboratory for Clinical and Experimental Physiology, Department of Medical Physiology, University of Vienna, Vienna, Austria
,
J C Kirchheimer
The Laboratory for Clinical and Experimental Physiology, Department of Medical Physiology, University of Vienna, Vienna, Austria
,
Liselotte Turcu
The Laboratory for Clinical and Experimental Physiology, Department of Medical Physiology, University of Vienna, Vienna, Austria
,
G Christ
The Laboratory for Clinical and Experimental Physiology, Department of Medical Physiology, University of Vienna, Vienna, Austria
,
B R Binder
The Laboratory for Clinical and Experimental Physiology, Department of Medical Physiology, University of Vienna, Vienna, Austria
› Author Affiliations
Further Information

Publication History

Received 21 January 1986

Accepted 25 March 1986

Publication Date:
18 July 2018 (online)

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Summary

Monoclonal antibodies against urinary urokinase were obtained by immunizing mice with purified human high molecular weight urokinase. Five antibodies were selected and denominated MPW1UK, MPW2UK, MPW3UK, MPW4UK, and MPW5UK, respectively. All selected antibodies reacted with high and low molecular weight urokinase. Cleavage of the low molecular weight paranitroanilide substrate pyro-Glu-Gly-Arg-pNA by urokinase was not inhibited by the antibodies and only one antibody (MPW5UK) inhibited plasminogen activation by urokinase. The ability of MPW5UK to bind to coated urokinase was 100-fold higher than that of the other antibodies. MPW5UK was used to prepare an immunosorbent for the purification of urokinase antigen from freshly voided crude urine. One-chain prourokinase was separated from two-chain urokinase by chromatography of the urokinase antigen containing mixture on agmatine Sepharose. As judged by SDS gel electrophoresis one-chain prourokinase as well as two-chain urokinase were purified to apparent homogeneity by this two-step procedure; the yields were 18% and 47% for single-chain prourokinase and two-chain urokinase, respectively, as calculated from total urokinase antigen contained in the starting material.

Keywords

Monoclonal antibodies - Two-chain urokinase - Single-chain prourokinase - Affinity purification

* Part of this paper was presented at the 7th International Congress on Fibrinolysis, Venice 1984, and published in abstract form (Haemostas 1984; 14: 43)


 

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