PURPOSE: To improve purity and yield of urokinase by adding a specific precipitating agent to human urine, eluting the precipitated urokinase with an EDTA solution and purifying the resultant elution with a benzidine sepharose affinity column.
CONSTITUTION: A base or an alkaline aqueous solution is added to human urine to adjust its pH to 7.5-8.0, the treated urine is allowed to stand to precipitate most solid components in the urine, and the resultant mixture is filtered to collect the supernatant solution. To the supernatant solution whose pH has been adjusted to 4-6 by adding an acid solution is added zinc chloride or copper sulfate as a precipitating agent in such an amount that the final concentration becomes 1-2 mM, and subsequently a sodium hydroxide aqueous solution to adjust pH to neutral to precipitate urokinase. Then, the resultant mixture containing the urokinase is centrifugally separated to collect the precipitated urokinase. The urokinase is eluted by dissolving the precipitated urokinase in a high concentration EDTA solution having pH of 7.0-8.0, and the eluted urokinase is purified by benzidine sepharose affinity column chromatography.
U KANSHIYOU
SHIN KOUJIYUN
HAKU SHIYOUFUKU
