Purification of High Molecular Weight Urokinase by Reverse-immunoadsorption

抄録

To develop a rapid and convenient purification procedure for high molecular weight urokinase (the native form, mol wt., 54, 000), a reverse-immunoadsorbent which binds only contaminating proteins in the crude urokinase preparation was synthesized. Essentially all the impurity proteins were removed by adsorption to the anti-contaminant-IgG Sepharose 4B without conversion of the high molecular weight urokinase to the low molecular weight urokinase; three to four-fold purification of the enzyme was achieved by this single procedure with a yield of about 90%. The homogeneous high molecular weight urokinase was obtained easily by a following elution from a CM-Sephadexcolumn. The overall purification of the enzyme took less than three days with a final yield of 88%. The ability of the reverse-immunoadsorbent to adsorb impurity proteins was not reduced after six batch reactions. Reverse-immunoadsorption may be a generally useful method for protein purification.