Abstract An initial solution of crude urokinase especially of human origin, is subjected to exclusion chromatography by contact with a DEAE cellulose resin, following adjustments of the pH of the solution to a value of from 4 to 6, and of its conductivity to a value of from 15,000 to 25,000 micromhos, and an effluent is collected to provide a preparation enriched in urokinase. A urokinase, mixed with foreign proteins, especially pyrogens, is purified by a partial saturation of an aqueous solution of this mixture by ammonium sulphate to a value for which the precipitate formed would not entrain more than 5% of the total activity of urokinase of the initial solution. The supernatent liquid is collected. A complex of urokinase and heparin, urokinase heparinate, is made by reacting a purified solution of urokinase with a solution of heparin. It has relative activities of 30,000 to 100,000 CTA units of urokinase for 100 Iu of heparin. In an isotonic perfusion solution in glucose serum, its dilution is from 300,000 to 900,000 CTA units of urokinase/250 ml.
A UROKINASE-HEPARINE COMPLEX COMPRISING HEPARINE COMPLEXED WITH HIGHLY PURIFIED UROKINASE, THE COMPLEX HAVING RETAINED ESSENTIALLY THE UROKINASE ACTIVITY.
Claim 2
Dependent Claim
18 words
A UROKINASE-HEPARINE COMPLEX COMPRISING HEPARINE COMPLEXED WITH HIGHLY PURIFIED UROKINASE, THE COMPLEX HAVING RETAINED ESSENTIALLY THE UROKINASE ACTIVITY.
Claim 3
Dependent Claim
21 words
The complex of claim 1 wherein the urokinase is a purified urokinase having a urokinase activity of at least 46,000 CTAu/mg.
Claim 4
Dependent Claim
18 words
The complex of claim 1 wherein the activity of heparine respective the activity of urokinase is comparatively negligible.
Claim 5
Dependent Claim
31 words
The complex of claim 1 wherein the ratio of heparine to urokinase activity is 100 Iu per mg of heparine to 30,000 to 100, 000 CTA units per mg or urokinase.
Claim 6
Dependent Claim
8 words
A lyophilizate of the complex of claim 1.
Claim 7
Dependent Claim
9 words
An anhydrous complex of the complex of claim 1.
Claim 8
Dependent Claim
9 words
An aqueous solution containing the lyophilizate of claim 5.
Claim 9
Independent Claim
44 words
The method of stabilizing urokinase which comprises reacting urokinase with heparin by contacting a solution of purified urokinase with a heparin solution to form a complex of purified urokinase and heparin, and recovering the complex, the complex having retained essentially the initial urokinase activity.
Claim 10
Independent Claim
44 words
The method of stabilizing urokinase which comprises reacting urokinase with heparin by contacting a solution of purified urokinase with a heparin solution to form a complex of purified urokinase and heparin, and recovering the complex, the complex having retained essentially the initial urokinase activity.
Claim 11
Dependent Claim
20 words
Method according to claim 8, wherein the reaction is effected at a pH in the range from 1 to 5.
Claim 12
Dependent Claim
18 words
Method according to claim 9, wherein the reaction is effected at a pH of the order of 4.2.
Claim 13
Dependent Claim
41 words
The method of claim 8 which comprises dissolving the complex in an aqueous solution, lyophilizing the solution, thereby obtaining an anhydrous complex. 12.. The method of claim 8 which comprises forming an aqueous solution of the complex and lyophilizing said solution.
Claim 14
Dependent Claim
12 words
The process of claim 8 which comprises separating the complex by precipitation.
Claim 15
Dependent Claim
15 words
The process of claim 8 which comprises precipitating the complex by acidifying this reaction medium.