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    Method of producing lys-plasminogen
    1.
    发明授权
    Method of producing lys-plasminogen 失效
    产生溶酶原纤溶酶原的方法

    公开(公告)号：US5371007A

    公开(公告)日：1994-12-06

    申请号：US41332

    申请日：1993-04-01

    申请人： Yendra Linnau ,  Ernst Hetzl

    发明人： Yendra Linnau ,  Ernst Hetzl

    IPC分类号： A61K38/46 ,  A61P7/02 ,  C12N9/68 ,  C12S3/14 ,  C12P21/00 ,  C12N9/00

    CPC分类号： C12N9/6435 ,  C12Y304/21007

    摘要： To produce lys-plasminogen having a specific activity of at least 17.5 caseinolytic units/mg protein and at least 50 mymoles/g protein nitrogen as well as an electrophoretic purity of at least 90%, plasminogen from plasma, a plasminogen-containing fraction or a tissue culture is adsorbed on immobilized lysine for the purpose of purification, is eluted and is recovered from the eluate by a protein precipitating agent. A solution of the thus purified plasminogen is adjusted to a plasmin activity ranging between 0.005 and 0.2 mymoles/ml min relative to chromogenic substrate H-D-valyl-L-leucyl-L-lysine-p-nitroanilide dihydrochloride, is maintained at a temperature of from +1.degree. C. to +20.degree. C. for a period of from 6 to 60 hours in order to provoke an enzymatic-proteolytic conversion of plasminogen into lys-plasminogen, whereupon the enzymatic action is interrupted and lys-plasminogen is isolated.

    摘要翻译： 为了产生具有至少17.5个酪蛋白分解单位/ mg蛋白和至少50 mymoles / g蛋白质氮的比活性以及至少90％的电泳纯度的溶血纤溶酶原，血浆中的纤溶酶原，含纤溶酶原的级分或 组织培养物被吸附在固定的赖氨酸上用于纯化，被洗脱并通过蛋白质沉淀剂从洗脱液中回收。 将如此纯化的纤溶酶原的溶液调节至相对于显色底物HD-缬氨酰-L-亮氨酰-L-赖氨酸 - 对硝基苯胺二盐酸盐在0.005至0.2 mymoles / ml min范围内的纤溶酶活性维持在 +1℃至+ 20℃，时间为6至60小时，以引起纤溶酶原转化为溶酶原纤维蛋白溶酶原的酶 - 蛋白水解转化，由此中断酶促作用并分离裂解纤溶酶原。

    Method of producing LYS-plasminogen
    3.
    发明授权
    Method of producing LYS-plasminogen 失效
    生产LYS-纤溶酶原的方法

    公开(公告)号：US5556766A

    公开(公告)日：1996-09-17

    申请号：US292290

    申请日：1994-08-18

    申请人： Yendra Linnau ,  Ernst Hetzl

    发明人： Yendra Linnau ,  Ernst Hetzl

    IPC分类号： A61K38/46 ,  A61P7/02 ,  C12N9/68 ,  C12S3/14 ,  C12P21/06 ,  C12N9/00

    CPC分类号： C12N9/6435 ,  C12Y304/21007

    摘要： A process for preparing lys-plasminogen from blood plasma or blood plasma products having a specific activity of at least 17.5 caseinolytic units/mg protein and at lease 50 .mu.moles/g protein nitrogen as well as a an electrophoretic purity of at least 90% is presented. Cohn fraction III is obtained by phosphate precipitation from blood plasma or blood plasma products. A crude plasminogen fraction is then obtained by precipitating most non-plasminogen proteins with ethanol. The crude plasminogen is absorbed on immobilized lysine affinity column. Elution followed by precipitation obtains a purified plasminogen. The purified plasminogen is converted to lys-plasminogen with plasmin having an activity from 0.01 to 0.1 .mu.moles/ml min relative to chromogenic substrate H-D-Val-L-Leu-L-Lys-p-nitroanilide by dialyzing at a temperature of about 1.degree. to 20.degree. C. for a time period of about 6 to 60 hours. The conversion is interrupted by adding aprotinin. The resulting lys-plasminogen is lyophilized to produce a lyophilized lys-plasminogen.

    摘要翻译： 从具有至少17.5％酪蛋白溶解单位/ mg蛋白质的比活性和至少50μg/ g蛋白质氮的比活性以及至少90％的电泳纯度的血浆或血浆产物制备lys-纤溶酶原的方法是 呈现。 通过从血浆或血浆产物中的磷酸盐沉淀获得Cohn级分III。 然后通过用乙醇沉淀大多数非纤溶酶原蛋白来获得粗纤溶酶原级分。 粗纤溶酶原被固定化的赖氨酸亲和柱吸收。 洗脱后沉淀得到纯化的纤溶酶原。 通过在约1℃的温度下透析，相对于显色底物HD-Val-L-Leu-L-Lys-对硝基苯胺将纯化的纤溶酶原转化为具有0.01至0.1μmol/ ml min活性的纤溶酶的lys-纤溶酶原 在-20℃下，时间约6〜60小时。 转化通过加入抑肽酶中断。 将所得的溶血纤维蛋白溶酶原冻干，以产生冻干溶胞纤溶酶原。