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公开(公告)号：US20070065906A1
公开(公告)日：2007-03-22
申请号：US10547802
申请日：2004-02-20
申请人： Satoshi Koyanagi , Keishin Sugawara , Yoshinobu Miyatsu , Toshio Murakami , Toshihiro Maeda , Hiroshi Mizokami
发明人： Satoshi Koyanagi , Keishin Sugawara , Yoshinobu Miyatsu , Toshio Murakami , Toshihiro Maeda , Hiroshi Mizokami
IPC分类号： C12P21/06 , C07H21/04 , C07K14/435 , C07K14/195 , C12N1/21 , C12N15/74
CPC分类号： C07K14/43531 , C12N15/70 , C12P21/02
摘要： A heterologous protein free from an inducer and a method for producing said protein are provided. A method for producing a heterologous protein which comprises the step of optionally culturing at low temperature recombinant E. coli cells expressing a heterologous protein under control of a promoter capable of inducing expression through temperature shift and then culturing at high temperature said recombinant E. coli cells in the absence of an inducer to thereby allow for expression of said heterologous protein, or the step of culturing at high temperature said recombinant E. coli cells to thereby simultaneously allow for both cell proliferation and expression of said heterologous protein, and the heterologous protein obtained by said method that is free from an inducer. Such heterologous protein may include a major mite allergen, a secretary macrophage toxin from Actinobacillus pleuropneumoniae and a surface protective antigen (SpaA) of Erysipelothrix rhusiopathiae.
摘要翻译： 提供了不含诱导物的异源蛋白质及其制备方法。 一种产生异源蛋白质的方法，其包括在低温下表达异源蛋白质的重组大肠杆菌细胞的步骤，所述重组大肠杆菌细胞在能够通过温度转移诱导表达并且然后在高温下培养的启动子控制下，所述重组大肠杆菌细胞 在不存在诱导剂从而允许所述异源蛋白质的表达的情况下，或在高温下培养所述重组大肠杆菌细胞从而同时允许所述异源蛋白质的细胞增殖和表达的步骤，获得的异源蛋白质 通过所述方法不含诱导剂。 这种异源蛋白可以包括主要的螨变应原，来自胸膜肺炎放线杆菌的秘密巨噬细胞毒素和Erysipelothrix rhusiopathiae的表面保护性抗原（SpaA）。
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公开(公告)号：US07667009B2
公开(公告)日：2010-02-23
申请号：US10555883
申请日：2004-04-14
申请人： Satoshi Koyanagi , Kenjiro Kawatsu , Toshio Murakami , Yoshinobu Miyatsu , Toshihiro Maeda , Hiroshi Mizokami
发明人： Satoshi Koyanagi , Kenjiro Kawatsu , Toshio Murakami , Yoshinobu Miyatsu , Toshihiro Maeda , Hiroshi Mizokami
IPC分类号： A23J1/00
CPC分类号： C07K14/43531 , A61K38/00 , A61K39/00
摘要： A method for purifying a modified major mite allergen obtained by the genetic recombination technique and a purified modified major mite allergen obtained by said method for purification are provided. A method for purifying a modified major mite allergen obtained by the genetic recombination technique, which comprises the purification steps: (1) Washing and recovering inclusion bodies containing a modified major mite allergen obtained by the genetic recombination technique with MF membrane; (2) Dissolving said inclusion bodies followed by refolding; (3) Concentrating a solution containing the modified major mite allergen with simultaneous removal of low molecular weight components with ultrafiltration membrane; (4) Recovering the modified major mite allergen in non-adsorbed fractions with an anion exchanger; (5) Recovering the modified major mite allergen in adsorbed fractions with a hydrophobic gel; and (6) Recovering the modified major mite allergen in adsorbed fractions with an anion exchanger, and a modified major mite allergen with high purity obtained by said method for purification.
摘要翻译： 提供了通过遗传重组技术获得的改良的主要螨变应原的纯化方法和通过所述纯化方法获得的纯化的改良的主要螨变应原。 一种用于纯化通过遗传重组技术获得的改性主要螨变应原的方法，其包括纯化步骤：（1）洗涤并回收含有通过MF膜的遗传重组技术获得的修饰的主要螨变应原的包含体; （2）溶解所述包涵体，然后重折叠; （3）浓缩含有改性主要螨变应原的溶液，同时用超滤膜除去低分子量成分; （4）用阴离子交换剂回收未吸附级分的改性主要螨变应原; （5）用疏水凝胶回收吸附级分中的改性主要螨变应原; 和（6）用阴离子交换剂回收吸附级分中的改性主要螨变应原，以及通过所述纯化方法获得的具有高纯度的改性主要螨变应原。
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公开(公告)号：US20080113424A1
公开(公告)日：2008-05-15
申请号：US10555883
申请日：2004-04-14
申请人： Satoshi Koyanagi , Kenjiro Kawatsu , Toshio Murakami , Yoshinobu Miyatsu , Toshihiro Maeda , Hiroshi Mizokami
发明人： Satoshi Koyanagi , Kenjiro Kawatsu , Toshio Murakami , Yoshinobu Miyatsu , Toshihiro Maeda , Hiroshi Mizokami
CPC分类号： C07K14/43531 , A61K38/00 , A61K39/00
摘要： A method for purifying a modified major mite allergen obtained by the genetic recombination technique and a purified modified major mite allergen obtained by said method for purification are provided. A method for purifying a modified major mite allergen obtained by the genetic recombination technique, which comprises the purification steps: (1) Washing and recovering inclusion bodies containing a modified major mite allergen obtained by the genetic recombination technique with MF membrane; (2) Dissolving said inclusion bodies followed by refolding; (3) Concentrating a solution containing the modified major mite allergen with simultaneous removal of low molecular weight components with ultrafiltration membrane; (4) Recovering the modified major mite allergen in non-adsorbed fractions with an anion exchanger; (5) Recovering the modified major mite allergen in adsorbed fractions with a hydrophobic gel; and (6) Recovering the modified major mite allergen in adsorbed fractions with an anion exchanger, and a modified major mite allergen with high purity obtained by said method for purification.
摘要翻译： 提供了通过遗传重组技术获得的改良的主要螨变应原的纯化方法和通过所述纯化方法获得的纯化的改良的主要螨变应原。 一种用于纯化通过遗传重组技术获得的改良的主要螨变应原的方法，其包括纯化步骤：（1）洗涤并回收含有通过MF膜的遗传重组技术获得的改性主要螨变应原的包含体; （2）溶解所述包涵体，然后重折叠; （3）浓缩含有改性主要螨变应原的溶液，同时用超滤膜除去低分子量成分; （4）用阴离子交换剂回收未吸附级分的改性主要螨变应原; （5）用疏水凝胶回收吸附级分中的改性主要螨变应原; 和（6）用阴离子交换剂回收吸附级分中的改性主要螨变应原，以及通过所述纯化方法获得的具有高纯度的改性主要螨变应原。
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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：US08332159B2
公开(公告)日：2012-12-11
申请号：US10546293
申请日：2004-02-18
IPC分类号： G06F7/00
CPC分类号： C07K16/00 , A61K2039/505 , C07K16/1063 , C07K2317/34
摘要： A method of enhancing the efficacy of a monoclonal antibody preparation wherein antigens from patients are tested for their reactivity with the antibody. An amino acid sequence of an expressed protein is deuced from a nucleotide sequence determined by isolation and analysis of a target molecule gene in a biopsy from a patient. This is compared with the previously determined amino acid sequence recognized by the monoclonal antibody preparation in order to assess the fitness of patients for administration of the monoclonal antibody preparation.
摘要翻译： 一种提高单克隆抗体制剂的功效的方法，其中测试来自患者的抗原与抗体的反应性。 通过从患者的活组织检查中分离和分析靶分子基因而确定的核苷酸序列，从表达的蛋白质的氨基酸序列中去除。 将其与由单克隆抗体制剂识别的先前确定的氨基酸序列进行比较，以评估患者施用单克隆抗体制剂的适合性。
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公开(公告)号：US20060280751A1
公开(公告)日：2006-12-14
申请号：US10546293
申请日：2004-02-18
CPC分类号： C07K16/00 , A61K2039/505 , C07K16/1063 , C07K2317/34
摘要： A method for enhancing efficacy of a monoclonal antibody preparation is provided wherein antigens from patients are tested for their reactivity with said antibody. In accordance with the method of the invention, an amino acid sequence of an expressed protein is deduced from a nucleotide sequence determined by isolation and analysis of a target molecule gene in biopsy from patients and is compared with the previously determined amino acid sequence recognized by said monoclonal antibody preparation to thereby assess fitness of patients for administration of said monoclonal antibody preparation.
摘要翻译： 提供了增强单克隆抗体制剂功效的方法，其中测试来自患者的抗原与其抗体的反应性。 根据本发明的方法，通过从患者的活组织检查中分离和分析靶分子基因而确定的核苷酸序列推导出表达蛋白质的氨基酸序列，并将其与先前确定的所述 从而评估患者适用于所述单克隆抗体制剂的施用。
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8.发明授权Method for preparing human serum albumin through heat-treatment in the presence of divalent cation 有权在二价阳离子存在下通过热处理制备人血清白蛋白的方法公开(公告)号：US07531631B2
公开(公告)日：2009-05-12
申请号：US11489541
申请日：2006-07-20
IPC分类号： C07K14/00
CPC分类号： C07K1/30 , C07K14/765
摘要： The present invention herein discloses a method for purifying human serum albumin, which is characterized in that it comprises the steps of heat-treating a human serum albumin-containing solution including impurities originated from a host cell, in the presence of a divalent cation such as calcium ion, magnesium ion, nickel ion, cobalt ion, iron ion and zinc ion to thus selectively allow the impurities to undergo agglutination. The present invention also provides the highly purified human serum albumin prepared by the foregoing method.
摘要翻译： 本发明的本发明公开了一种纯化人血清白蛋白的方法，其特征在于，其包括在二价阳离子如二氯甲烷存在下热处理含有源自宿主细胞的杂质的人血清白蛋白溶液的步骤 钙离子，镁离子，镍离子，钴离子，铁离子和锌离子，从而选择性地允许杂质发生凝集。 本发明还提供通过上述方法制备的高纯度人血清白蛋白。
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9.发明申请公开(公告)号：US20060258850A1
公开(公告)日：2006-11-16
申请号：US11489541
申请日：2006-07-20
IPC分类号： C12P21/06 , C07H21/04 , C07K14/765
CPC分类号： C07K1/30 , C07K14/765
摘要： The present invention herein discloses a method for purifying human serum albumin, which is characterized in that it comprises the steps of heat-treating a human serum albumin-containing solution including impurities originated from a host cell, in the presence of a divalent cation such as calcium ion, magnesium ion, nickel ion, cobalt ion, iron ion and zinc ion to thus selectively allow the impurities to undergo agglutination. The present invention also provides the highly purified human serum albumin prepared by the foregoing method.
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公开(公告)号：US07001885B2
公开(公告)日：2006-02-21
申请号：US10175382
申请日：2002-06-20
申请人： Satoshi Adachi , Hiroshi Mizokami , Yoshitaka Tajima , Yoshinobu Miyatsu , Toshinobu Nouchi , Yoshitaka Ushio
发明人： Satoshi Adachi , Hiroshi Mizokami , Yoshitaka Tajima , Yoshinobu Miyatsu , Toshinobu Nouchi , Yoshitaka Ushio
IPC分类号： A61K38/00
CPC分类号： C07K14/765 , C07K1/18
摘要： Multimers of human serum albumin are removed from a solution of human serum albumin containing the multimers by bringing the human serum albumin solution into contact with an anion-exchanger equilibrated with a buffer containing a salt in a concentration ranging from 10 to 150 mM and having a pH value ranging from 5 to 9.5.
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