Wojta J, Kirchheimer J C, Turcu L, Christ G, Binder B R
Thromb Haemost. 1986 Jun 30;55(3):347-51.
Monoclonal antibodies against urinary urokinase were obtained by immunizing mice with purified human high molecular weight urokinase. Five antibodies were selected and denominated MPW1UK, MPW2UK, MPW3UK, MPW4UK, and MPW5UK, respectively. All selected antibodies reacted with high and low molecular weight urokinase. Cleavage of the low molecular weight paranitroanilide substrate pyro-Glu-Gly-Arg-pNA by urokinase was not inhibited by the antibodies and only one antibody (MPW5UK) inhibited plasminogen activation by urokinase. The ability of MPW5UK to bind to coated urokinase was 100-fold higher than that of the other antibodies. MPW5UK was used to prepare an immunosorbent for the purification of urokinase antigen from freshly voided crude urine. One-chain prourokinase was separated from two-chain urokinase by chromatography of the urokinase antigen containing mixture on agmatine Sepharose. As judged by SDS gel electrophoresis one-chain prourokinase as well as two-chain urokinase were purified to apparent homogeneity by this two-step procedure; the yields were 18% and 47% for single-chain prourokinase and two-chain urokinase, respectively, as calculated from total urokinase antigen contained in the starting material.
通过用纯化的人高分子量尿激酶免疫小鼠，获得了抗尿中尿激酶的单克隆抗体。选择了五种抗体，分别命名为MPW1UK、MPW2UK、MPW3UK、MPW4UK和MPW5UK。所有选定的抗体均与高分子量和低分子量尿激酶发生反应。尿激酶对低分子量对硝基苯胺底物焦谷氨酸-甘氨酸-精氨酸-pNA的切割未被这些抗体抑制，只有一种抗体（MPW5UK）抑制尿激酶对纤溶酶原的激活。MPW5UK与包被的尿激酶结合的能力比其他抗体高100倍。MPW5UK用于制备免疫吸附剂，以从新鲜排出的粗尿中纯化尿激酶抗原。通过将含尿激酶抗原的混合物在胍丁胺琼脂糖上进行层析，将单链尿激酶原与双链尿激酶分离。通过SDS凝胶电泳判断，通过这两步程序，单链尿激酶原和双链尿激酶均被纯化至表观均一性；根据起始材料中所含总尿激酶抗原计算，单链尿激酶原和双链尿激酶的产率分别为18%和47%。
