Andrassy K, Ritz E
Thromb Haemost. 1977 Aug 31;38(2):536-44.
Urokinase was highly purified by electrophoretical and immunological methods starting with a commercial urokinase preparation (UK-Leo). Contaminating serum proteins and enzyme activities migrated into opposite directions in agar gel electrophoresis which proved to be a valuable preparative method. The final purification achieved was 80,000 Ploug units/mg protein. Traces of albumin, alpha2HS-glycoprotein and alpha2-macroglobulin migrated towards the cathode together with UK in a multimolecular complex. Urokinase antibodies (rabbit) gave with the cathodic fraction 2 precipitation lines (Ouchterlony technique): the one precipitation line corresponded to urokinase (molecular weight on gel chromatography 32,000 daltons), the other corresponded to UK complexed with serum proteins. Urokinase antibodies completely suppressed UK activity in various commercial preparations. All these preparations showed immunological identity; on disc electrophoresis pure urokinase (32,000 daltons, 80,000 Ploug units/mg protein) still gave 2--3 bands suggesting the presence of isoenzymes.
以市售尿激酶制剂（UK - Leo）为起始原料，通过电泳和免疫学方法对尿激酶进行了高度纯化。在琼脂凝胶电泳中，污染的血清蛋白和酶活性向相反方向迁移，这被证明是一种有价值的制备方法。最终实现的纯化程度为80,000普洛格单位/毫克蛋白质。微量的白蛋白、α2HS - 糖蛋白和α2 - 巨球蛋白与尿激酶一起以多分子复合物的形式向阴极迁移。尿激酶抗体（兔源）与阴极部分产生2条沉淀线（欧氏免疫双扩散技术）：一条沉淀线对应尿激酶（凝胶色谱法测定分子量为32,000道尔顿），另一条对应与血清蛋白复合的尿激酶。尿激酶抗体完全抑制了各种市售制剂中的尿激酶活性。所有这些制剂都显示出免疫同一性；在圆盘电泳中，纯尿激酶（32,000道尔顿，80,000普洛格单位/毫克蛋白质）仍出现2 - 3条带，表明存在同工酶。
