RAPID PURIFICATION OF STREPTOKINASE BY AFFINITY CHROMATOGRAPHY

Affinity columns prepared with partially degraded plasmin provide a means of rapidly purifying streptokinase from crude preparations. Human plasmin was formed from plasminogen by incubation with small amounts of streptokinase at 37°C. The plasmin was held at this temperature for 2 h; extensive degradation of the light chain of plasmin resulted and little plasmin activity remained after this period. Any remaining active plasmin was inhibited with diisopropylfluorophosphate. The plasmin was then bound to agarose. Streptokinase was selectively absorbed from a crude preparation and relatively mild conditions were used to elute the streptokinase from the column. More than 93% of the starting streptokinase activity was recovered with a 5.0 fold increase in specific activity. The streptokinase produced had a single NH2-terminal amino acid sequence identical to that of standard streptokinase preparations. No evidence of contamination by plasmin was found. © 1979.