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1.发明专利
公开(公告)号：JP2014064594A
公开(公告)日：2014-04-17
申请号：JP2014007656
申请日：2014-01-20
发明人： COX GEORGE N , DOHERTY DANIEL H , ROSENDAHL MARY S
IPC分类号： C12N15/09 , C12P21/02 , A61K38/04 , A61K38/21 , A61K38/22 , A61K38/27 , A61K47/48 , A61P5/02 , A61P7/06 , A61P43/00 , C07K14/47 , C07K14/505 , C07K14/56 , C07K14/575 , C07K14/61 , C07K14/72 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To provide a method of obtaining soluble proteins having at least one added free cysteines.SOLUTION: The method comprises the steps of: (a) obtaining a host cell capable of expressing the soluble protein having at least one added free cysteines; (b) exposing the host cell to a cysteine blocking agent forming a stable mixed disulfide with the at least one added free cysteines before the step (c); and (c) isolating the soluble protein from the host cell. The produced soluble proteins can be modified to increase their effectiveness, and such modifications include attaching a PEG moiety to form pegylated proteins.
摘要翻译： 要解决的问题：提供一种获得具有至少一种添加的游离半胱氨酸的可溶性蛋白质的方法。溶液：该方法包括以下步骤：（a）获得能够表达具有至少一种添加的游离半胱氨酸的可溶性蛋白质的宿主细胞 ; （b）在步骤（c）之前将所述宿主细胞暴露于形成稳定的混合二硫化物的半胱氨酸阻断剂与所述至少一种加入的游离半胱氨酸; 和（c）从宿主细胞中分离可溶性蛋白质。 产生的可溶性蛋白质可以被修饰以提高它们的有效性，并且这些修饰包括连接PEG部分以形成聚乙二醇化蛋白质。
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公开(公告)号：JP2011173911A
公开(公告)日：2011-09-08
申请号：JP2011098711
申请日：2011-04-26
发明人： ROSENDAHL MARY S , COX GEORGE N , DOHERTY DANIEL H
IPC分类号： C07K14/535 , C12N15/09 , C07K1/02 , C07K1/107 , C07K1/113 , C07K14/00 , C07K14/505 , C07K14/515 , C07K14/52 , C07K14/56 , C07K14/61 , C07K14/78 , C12P21/02
CPC分类号： C07K14/52 , A61K47/60 , C07K1/1133 , C07K14/505 , C07K14/515 , C07K14/535 , C07K14/56 , C07K14/61 , C07K14/78
摘要： PROBLEM TO BE SOLVED: To provide novel methods for making and refolding insoluble or aggregated proteins having free cysteine. SOLUTION: The present invention relates to novel methods for making and refolding insoluble or aggregated proteins having free cysteines in which a host cell expressing the protein is exposed to a cysteine blocking agent. The soluble, refolded proteins produced by the novel methods can then be modified to increase their effectiveness. Such modifications include attaching a PEG moiety to form PEGylated proteins. COPYRIGHT: (C)2011,JPO&INPIT
摘要翻译： 要解决的问题：提供用于制备和重折叠具有游离半胱氨酸的不溶性或聚集蛋白质的新方法。 解决方案：本发明涉及制备和重折叠具有游离半胱氨酸的不溶性或聚集蛋白质的新方法，其中表达蛋白质的宿主细胞暴露于半胱氨酸阻断剂。 然后可以修饰通过新方法产生的可溶的，重折叠的蛋白质以增加它们的有效性。 这样的修饰包括连接PEG部分以形成聚乙二醇化蛋白质。 版权所有（C）2011，JPO＆INPIT
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公开(公告)号：JP2010280714A
公开(公告)日：2010-12-16
申请号：JP2010196790
申请日：2010-09-02
发明人： COX GEORGE N , DOHERTY DANIEL H , ROSENDAHL MARY S
IPC分类号： C07K14/61 , C12N15/09 , A61K38/04 , A61K38/21 , A61K38/22 , A61K38/27 , A61K47/34 , A61K47/48 , A61P5/02 , A61P7/06 , A61P43/00 , C07K14/47 , C07K14/505 , C07K14/56 , C07K14/575 , C07K14/72 , C12P21/02 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To provide an isolated monoPEGylated growth hormone protein. SOLUTION: The isolated monoPEGylated growth hormone protein comprises a polyethylene glycol attached to an amino acid selected from the group consisting of amino acid 2, 3, 5, 38, 40, 55, 57, 99, 101, 102, 103, 132, 133, 134, 137, 140, 141, 143, 144, 147, 148, 149, 186, and 187 of growth hormone, wherein the obtained monoPEGylated growth hormone protein stimulates proliferation in vitro of a cell line that proliferates in response to growth hormone. COPYRIGHT: (C)2011,JPO&INPIT
摘要翻译： 待解决的问题：提供分离的单PEG化生长激素蛋白。 解决方案：分离的单PEG化生长激素蛋白包含与氨基酸相连的聚乙二醇，所述氨基酸选自氨基酸2,3,5,38,40,55,57,99,101,102,103,103， 生长激素的第132,133,134,137,140,141,143,144,147,148,149,186和187，其中所获得的单PEG化生长激素蛋白刺激体外增殖，所述细胞系响应于 生长激素。 版权所有（C）2011，JPO＆INPIT
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公开(公告)号：JP2009096810A
公开(公告)日：2009-05-07
申请号：JP2008302460
申请日：2008-11-27
发明人： COX GEORGE N III
IPC分类号： C07K14/565 , C12N15/09 , A61K38/21 , A61K47/48 , A61P5/10 , A61P43/00 , C07K14/475 , C07K14/505 , C07K14/52 , C07K14/53 , C07K14/535 , C07K14/54 , C07K14/55 , C07K14/555 , C07K14/56 , C07K14/57 , C07K14/61 , C07K17/08 , C12N15/12
CPC分类号： C07K14/5431 , A61K38/00 , A61K47/60 , C07K14/475 , C07K14/505 , C07K14/52 , C07K14/535 , C07K14/5403 , C07K14/5406 , C07K14/5409 , C07K14/5412 , C07K14/5418 , C07K14/5425 , C07K14/5428 , C07K14/5434 , C07K14/5437 , C07K14/5443 , C07K14/55 , C07K14/555 , C07K14/56 , C07K14/565 , C07K14/57 , C07K14/61
摘要： PROBLEM TO BE SOLVED: To provide a preferred site for adding a cysteine residue to the growth hormone supergene family comprising greater than 20 structurally related cytokine and growth factors, or for introducing cysteine substitution, and a protein and protein derivatives produced thereby. SOLUTION: A general method for preparing a site-specific biologically active conjugate of the growth hormone supergene family proteins such as β-interferon or the like is provided. This method comprises a step in which a cysteine residue is added to a non-essential region of this protein, or a step in which a non-essential amino acid in the protein is substituted with a cysteine residue by using site-specific mutagenesis, and then a step in which the protein is connected to a cysteine reactive polymer through the added cysteine residue or another type cysteine reactive part is connected with a covalent bond. COPYRIGHT: (C)2009,JPO&INPIT
摘要翻译： 要解决的问题：提供一个优选的位点，用于向包含大于20个结构相关的细胞因子和生长因子或用于引入半胱氨酸取代的生长激素超基因家族添加半胱氨酸残基，以及由此产生的蛋白质和蛋白质衍生物。 提供了制备生长激素超基因家族蛋白如β-干扰素等的位点特异性生物活性缀合物的一般方法。 该方法包括将半胱氨酸残基添加到该蛋白质的非必需区域的步骤，或通过使用位点特异性诱变将蛋白质中的非必需氨基酸置换为半胱氨酸残基的步骤，以及 那么蛋白质通过加入的半胱氨酸残基或另一类型的半胱氨酸反应性部分与半胱氨酸反应性聚合物连接的步骤与共价键连接。 版权所有（C）2009，JPO＆INPIT
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