第8卷‧第5期,
198005
, pp. 393-405
利用親和性柱層層析法精製尿激?之研究
- 作者:
蘇遠志; 杜錦治; 江晃榮
- 作者服務機構:
國立臺灣大學農業化學系
- 中文摘要:
尿激?是一種胞漿素原活化酵素(plasminogen activating enzyme),可以將血液中的胞漿
素原活化後變成胞漿素(plasmin),藉以分解血管中形成的血纖維凝塊(fibrin clot),以治療各
種血栓症,為世界各國十分重視的一種醫療藥劑。本實驗係利用親和性柱層層析法,將粗製尿激
?加以純化。
首先將粗酵素(比活性約為5,000~10,000 IU∕mg protein)經過DEAE-cellulose, Sepha-
dex G-100或CM-Sephadex C-50等前處理去除大部分的雜質及著色物質後,再通入合成的
親和性膠體Sepharose 4B-6-aminohexanoic acid-p-aminobenzamidine柱層,溶出後可以得到
純度很高的尿激?,其比活性高達80,OOO IU/mg protein以上,zero coagulant activity為
150 IU/ml plasma,並以15,000 IU/kg家?的熱原試驗結果,體溫的上升亦符合美國藥典的規
格,由Sephadex G-100的分子量測定得知分子量50,000的尿激?部分超過85%,已達現行
國際醫藥品級精製尿激?的規格。木純化方法對原料粗酵素具有50%以上的活性同收率。e
- 英文摘要:
Urokinase, a plasminogen activating pro-teolytic enzyme, has been used clinically for thetreatment of thrombolisms by the action of con-verting plasminogen to a proteolytic enzymeplasmin to dissolve the fibrin clot in blood. A rapid and economically feasible purifica-tion of urokinase was achieved by an affinitycolumn chromatography. The crude urokinase was pretreated withDEAF-cellulose, Sephadex G-100 or CM-Sephadex C-50 to remove a major part of impa-rities and the coloring materials and then passedthrough the affinity gel column of Sepharose4B-6-aminohexanoic acid-p-aminobenzamidine.Through this process, the purified urokinasewith following quality was obtained: specificactivity not lower than 80,000 IU/mg protein;zero coagulant activity 150 IU/ml plasma; passthe pyrogen test with 15,000 IU/kg rabbit. Themolecular weight of the major moieties of pu-rified urokinase was estimated to be 50,000 andover 85% of the total activity. Above mentionedquality coincided with the international speci-fication of clinical urokinase. The overall re-covery of activity from crude urokinase wasabout 50%.
- 中文關鍵字:
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- 英文關鍵字:
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