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公开(公告)号：US06869934B2
公开(公告)日：2005-03-22
申请号：US10088588
申请日：2001-07-18
申请人： Hiroshi Mizokami , Shinichi Furukawa , Keishin Sugawara , Tatsufumi Onchi , Kazuhiro Komatsu , Satoshi Koyanagi , Hideo Yoshizaki
发明人： Hiroshi Mizokami , Shinichi Furukawa , Keishin Sugawara , Tatsufumi Onchi , Kazuhiro Komatsu , Satoshi Koyanagi , Hideo Yoshizaki
CPC分类号： C12N9/6432 , C07K14/4721 , C07K14/4728 , C12Y304/21006
摘要： The present invention relates to a method for purifying a calcium ion-binding protein by cation exchange chromatography. The present invention provide a method for isolating and purifying a calcium ion-binding protein in a simple and efficient manner from a liquid sample containing a calcium ion-binding protein and contaminants without any pretreatment such as addition of a chelating agent. More specifically, the present invention relates to a method for purifying a calcium ion-binding protein which comprises contacting said protein with a cation exchange carrier in the presence of calcium ions to let the said protein be adsorbed to the carrier, and after washing, eluting said protein, and to a calcium ion-binding protein having substantially no contaminants obtained by the method of the present invention.
摘要翻译： 本发明涉及通过阳离子交换层析纯化钙离子结合蛋白的方法。 本发明提供了从含有钙离子结合蛋白质和污染物的液体样品中以简单有效的方式分离和纯化钙离子结合蛋白的方法，而没有任何预处理如加入螯合剂。 更具体地，本发明涉及一种纯化钙离子结合蛋白的方法，其包括在钙离子存在下使所述蛋白质与阳离子交换载体接触以使所述蛋白质吸附到载体上，并且在洗涤后，洗脱 所述蛋白质和基本上不含有通过本发明的方法获得的污染物的钙离子结合蛋白质。
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公开(公告)号：US20050059805A1
公开(公告)日：2005-03-17
申请号：US10944810
申请日：2004-09-21
申请人： Hiroshi Mizokami , Shinichi Furukawa , Keishin Sugawara , Tatsufumi Onchi , Kazuhiro Komatsu , Satoshi Koyanagi , Hideo Yoshizaki
发明人： Hiroshi Mizokami , Shinichi Furukawa , Keishin Sugawara , Tatsufumi Onchi , Kazuhiro Komatsu , Satoshi Koyanagi , Hideo Yoshizaki
CPC分类号： C12N9/6432 , C07K14/4721 , C07K14/4728 , C12Y304/21006
摘要： The present invention relates to a method for purifying a calcium ion-binding protein by cation exchange chromatography. The present invention provide a method for isolating and purifying a calcium ion-binding protein in a simple and efficient manner from a liquid sample containing a calcium ion-binding protein and contaminants without any pretreatment such as addition of a chelating agent. More specifically, the present invention relates to a method for purifying a calcium ion-binding protein which comprises contacting said protein with a cation exchange carrier in the presence of calcium ions to let the said protein be adsorbed to the carrier, and after washing, eluting said protein, and to a calcium ion-binding protein having substantially no contaminants obtained by the method of the present invention.
摘要翻译： 本发明涉及通过阳离子交换层析纯化钙离子结合蛋白的方法。 本发明提供了从含有钙离子结合蛋白质和污染物的液体样品中以简单有效的方式分离和纯化钙离子结合蛋白的方法，而没有任何预处理如加入螯合剂。 更具体地，本发明涉及一种纯化钙离子结合蛋白的方法，其包括在钙离子存在下使所述蛋白质与阳离子交换载体接触以使所述蛋白质吸附到载体上，并且在洗涤后，洗脱 所述蛋白质和基本上不含有通过本发明的方法获得的污染物的钙离子结合蛋白质。
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公开(公告)号：US20090137001A1
公开(公告)日：2009-05-28
申请号：US12088823
申请日：2006-09-29
IPC分类号： C12P21/02 , C12N9/74 , C07K14/745 , C12N5/06
CPC分类号： C12N9/6429 , C12Y304/21005
摘要： The present invention provides for a process for preparing a recombinant human thrombin. A process for preparing a recombinant human thrombin which comprises: (1) obtaining a transfectant cell producing a human prothrombin by introducing an expression vector, wherein a gene fragment coding for a human prothrombin gene is incorporated, into an animal cell; (2) purifying a human prothrombin from the culture of the transfectant cell above by an anion exchanger; (3) converting the purified human prothrombin into a human thrombin by subjecting said human prothrombin to the action of ecarin; and (4) purifying the human thrombin from the solution after treatment with ecarin by an affinity method using benzamidine and a cation exchanger, and human thrombin obtained by said process, and a CHO cell that produces human prothrombin.
摘要翻译： 本发明提供了制备重组人凝血酶的方法。 一种制备重组人凝血酶的方法，其包括：（1）通过将编码人凝血酶原基因的基因片段引入到动物细胞中的表达载体获得产生人凝血酶原的转染细胞; （2）通过阴离子交换剂从上述转染细胞的培养物纯化人凝血酶原; （3）通过使所述人凝血酶原受到ecarin的作用，将纯化的人凝血酶原转化为人凝血酶; 和（4）使用苯甲脒和阳离子交换剂的亲和法，通过所述方法获得的人凝血酶和产生人凝血酶原的CHO细胞，用ecarin处理后，从溶液中纯化人凝血酶。
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