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Thromb Haemost 1975; 34(01): 194-209
DOI: 10.1055/s-0038-1651367
Original Article
Schattauer GmbH

Purification and Some Properties of Urokinase

Nobuhisa Ogawa
1   Research Laboratory Mochida Pharmaceutical Company Limited, 1-1, Kamiya 1-chome, Kitaku, Tokyo, 115 Japan
,
Hiroshi Yamamoto
1   Research Laboratory Mochida Pharmaceutical Company Limited, 1-1, Kamiya 1-chome, Kitaku, Tokyo, 115 Japan
,
Tomoaki Katamine
1   Research Laboratory Mochida Pharmaceutical Company Limited, 1-1, Kamiya 1-chome, Kitaku, Tokyo, 115 Japan
,
Hideo Tajima
1   Research Laboratory Mochida Pharmaceutical Company Limited, 1-1, Kamiya 1-chome, Kitaku, Tokyo, 115 Japan
› Author Affiliations
Further Information

Publication History

Received 21 January 1975

Accepted 12 March 1975

Publication Date:
02 July 2018 (online)

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Summary

A method for efficient extraction of urokinase from human urine was established by using polyacrylonitrile synthetic fiber as an adsorbent. By a combination of this method and known methods for purification of proteins, such as gel filtration and ion-exchange chromatography, urokinase with a specific activity of 224,000 International Units per mg of protein was obtained.

This sample showed homogeneity by ultracentrifugation, moving-boundary electrophoresis at pH 4.8 and 9.0 and Polyacrylamide gel disc electrophoresis at pH 4.0, but was separated into five active fractions by isoelectric focusing and Polyacrylamide gel disc electrophoresis at pH 9.4. This sample showed a single precipitin line in double radial immunodiffusion and Immunoelectrophoresis using rabbit anti-urokinase serum. This precipitin line fused with that of the International Standard preparation of urokinase and its immunological identity was established.

The molecular weight of this sample was 33,000, agreeing with that of the International Standard preparation. Its optimal pH as a plasminogen activator was approximately 8.8.

 

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