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1. 发明申请

WO2004020611A1 MYROTHECIUM SP. TRANSFORMATION AND EXPRESSION SYSTEM 审中-公开
标题翻译： MYROTHECIUM SP。转换和表达系统
公开(公告)号：WO2004020611A1
公开(公告)日：2004-03-11
申请号：PCT/BE2003/000143
申请日：2003-08-29
申请人： PURATOS NAAMLOZE VENNOOTSCHAP , JONNIAUX, Jean-Luc , VALEPYN, Emmanuel , CORBISIER, Anne-Marie , DAUVRIN, Thierry
发明人： JONNIAUX, Jean-Luc , VALEPYN, Emmanuel , CORBISIER, Anne-Marie , DAUVRIN, Thierry
IPC分类号： C12N1/14
CPC分类号： C07K14/37 , C12N1/14 , C12N15/80
摘要： The present invention is related to a transformation and an expression system in which Myrothecium sp. host cells are used to express homologous or heterologous proteins or are used to genetically engineer metabolic pathways.
摘要翻译：本发明涉及一种转化和表达系统，其中Myrothecium sp。宿主细胞用于表达同源或异源蛋白质，或用于遗传工程代谢途径。

2. 发明申请

WO2009095235A1 METHODS TO STABILIZE PROTEINS AND POLYPEPTIDES 审中-公开
标题翻译：稳定蛋白质和多肽的方法
公开(公告)号：WO2009095235A1
公开(公告)日：2009-08-06
申请号：PCT/EP2009/000573
申请日：2009-01-29
申请人： ABLYNX N.V. , JONNIAUX, Jean-Luc , LAUWEREYS, Marc, Jozef , STANSSENS, Patrick
发明人： JONNIAUX, Jean-Luc , LAUWEREYS, Marc, Jozef , STANSSENS, Patrick
IPC分类号： C07K14/00 , C07K16/00
CPC分类号： A61K39/39591 , C07K16/36 , C07K2317/22 , C07K2317/92
摘要： Methods of modifying and in particular stabilizing proteins and polypeptides by which a predetermined amino acid is introduced into selected positions of said protein or polypeptide to produce a small group of mutants. The methods are based on the premise that certain amino acids play a crucial role in the stability of proteins or polypeptides. Generated mutants can then be further analysed for stability and/or function, e.g. affinity. Furthermore, appropriate mutants may be combined to result in further optimized proteins or polypeptides. In addition, stabilized example polypeptides and suitable methods to identify and/or analyse de-stabilized or stabilized proteins or polypeptides are provided. The methods can be used to study the role of specific amino acids in protein structure and function and to develop new or improved, e.g. stabilized proteins and polypeptides such as antibodies and single variable domains.
摘要翻译：修饰和特别是稳定蛋白质和多肽的方法，通过该蛋白质和多肽将预定的氨基酸引入所述蛋白质或多肽的选定位置以产生少量的突变体。这些方法基于某些氨基酸在蛋白质或多肽的稳定性中发挥关键作用的前提。然后可以进一步分析产生的突变体的稳定性和/或功能，例如。亲和力。此外，合适的突变体可以组合以产生进一步优化的蛋白质或多肽。此外，提供了稳定的实施例多肽和鉴定和/或分析去稳定化或稳定的蛋白质或多肽的合适方法。该方法可用于研究特定氨基酸在蛋白质结构和功能中的作用，并开发新的或改进的，例如，稳定的蛋白质和多肽，例如抗体和单可变结构域。

3. 发明申请

WO2010125187A3 METHOD FOR THE PRODUCTION OF DOMAIN ANTIBODIES 审中-公开
标题翻译：产生抗体的方法
公开(公告)号：WO2010125187A3
公开(公告)日：2011-01-20
申请号：PCT/EP2010055916
申请日：2010-04-30
申请人： ABLYNX NV , SCHOTTE PETER , STANSSENS PATRICK , LABEUR CHRISTINE , JONNIAUX JEAN-LUC , LAUWEREYS MARC JOZEF
发明人： SCHOTTE PETER , STANSSENS PATRICK , LABEUR CHRISTINE , JONNIAUX JEAN-LUC
IPC分类号： C07K16/00 , C07K16/18 , C07K16/28 , C12N15/81 , C12P1/02
CPC分类号： C12P21/005 , C07K16/00 , C07K16/18 , C07K16/2875 , C07K2317/14 , C07K2317/24 , C07K2317/35 , C07K2317/56 , C07K2317/569 , C07K2317/624 , C07K2317/626
摘要： The present disclosure relates to a method for producing a domain antibody in a host other than E. coli, preferably yeast, comprising a) applying conditions that promote the formation of disulfide bridges in domain antibodies, or b) removing domain antibodies lacking at least one disulfide bridge, or c) a combination of (a) and (b). More specifically, the present disclosure relates to a method, wherein said conditions that promote the formation of disulfide brides are selected from one or more of the following: a) addition of oxidizing agents, preferably oxidizing metal ions, preferably one or more selected from Cu2+, Fe2+, Fe3+ and Zn2+; b) co-expression of the domain antibody with a thiol lsomerase,- c) adapting the culturing conditions by one or more selected from the following: lowering culturing temperature and/or optimizing the culturing medium, including but not limited to reduction of methanol feed for hosts requiring a methanol feed, lowering conductivity of the culture medium, addition of yeast extract and/or peptone, or any combination thereof; d) refolding the domain antibody in the presence of denaturant and redox-buffer e) treating the domain antibody by oxygenation, increasing temperature, increasing pH, high pressure or any combination thereof, and f ) combinations of any of a) through e), or wherein conditions that remove domain antibodies lacking at least one disulfide bridge are selected from a) binding domain antibodies comprising free thiol groups to suitable reactive groups, including but not limited to immobilized thiol groups, optionally under denaturing conditions; b) reverse phase high performance chromatography.
摘要翻译：本公开内容涉及在除大肠杆菌以外，优选酵母中的宿主中产生结构域抗体的方法，其包括a）施加促进结构域抗体中二硫键形成的条件，或b）除去缺少至少一个二硫键，或c）（a）和（b）的组合。更具体地，本公开涉及一种方法，其中促进形成二硫化物新娘的所述条件选自以下一种或多种：a）加入氧化剂，优选氧化金属离子，优选选自Cu 2+的一种或多种，，Fe2 +，Fe3 +和Zn2 +; b）结构域抗体与硫醇异构酶共表达，c）使培养条件适应一种或多种选自以下的培养条件：降低培养温度和/或优化培养基，包括但不限于还原甲醇进料对于需要甲醇进料的主体，降低培养基的导电性，添加酵母提取物和/或蛋白胨或其任何组合; d）在变性剂和氧化还原缓冲剂存在下重折叠结构域抗体e）通过氧合，升高温度，增加pH，高压或其任何组合来处理结构域抗体，以及f）a）至e），或者其中除去缺少至少一个二硫键的结构域抗体的条件选自a）包含游离硫醇基团的结合域抗体适合于适当的反应性基团，包括但不限于固定硫醇基团，任选地在变性条件下; b）反相高效液相色谱法。

4. 发明申请

WO2010125187A2 METHOD FOR THE PRODUCTION OF DOMAIN ANTIBODIES 审中-公开
标题翻译：产生抗体的方法
公开(公告)号：WO2010125187A2
公开(公告)日：2010-11-04
申请号：PCT/EP2010/055916
申请日：2010-04-30
申请人： ABLYNX NV , SCHOTTE, Peter , STANSSENS, Patrick , LABEUR, Christine , JONNIAUX, Jean-Luc , LAUWEREYS, Marc Jozef
发明人： SCHOTTE, Peter , STANSSENS, Patrick , LABEUR, Christine , JONNIAUX, Jean-Luc
IPC分类号： C07K16/00 , C12N15/81 , C12P1/02 , C07K16/28 , C07K16/18
CPC分类号： C12P21/005 , C07K16/00 , C07K16/18 , C07K16/2875 , C07K2317/14 , C07K2317/24 , C07K2317/35 , C07K2317/56 , C07K2317/569 , C07K2317/624 , C07K2317/626
摘要： The present invention relates to a method for producing a domain antibody in a host other than E. coli, preferably yeast, comprising a) applying conditions that promote the formation of disulfide bridges in domain antibodies, or b) removing domain antibodies lacking at least one disulfide bridge, or c) a combination of (a) and (b). More specifically, the present invention relates to a method, wherein said conditions that promote the formation of disulfide brides are selected from one or more of the following: a) addition of oxidizing agents, preferably oxidizing metal ions, preferably one or more selected from Cu2+, Fe2+, Fe3+ and Zn2+; b) co-expression of the domain antibody with a thiol isomerase; c) adapting the culturing conditions by one or more selected from the following: lowering culturing temperature and/or optimizing the culturing medium, including but not limited to reduction of methanol feed for hosts requiring a methanol feed, lowering conductivity of the culture medium, addition of yeast extract and/or peptone, or any combination thereof; d) refolding the domain antibody in the presence of denaturant and redox-buffer e) treating the domain antibody by oxygenation, increasing temperature, increasing pH, high pressure or any combination thereof, and f ) combinations of any of a) through e), or wherein conditions that remove domain antibodies lacking at least one disulfide bridge are selected from a) binding domain antibodies comprising free thiol groups to suitable reactive groups, including but not limited to immobilized thiol groups, optionally under denaturing conditions; b) reverse phase high performance chromatography.
摘要翻译：本发明涉及在除大肠杆菌以外，优选酵母中的宿主中产生结构域抗体的方法，该方法包括：a）施加促进结构域抗体中二硫键形成的条件，或b）去除缺少至少一个二硫键，或c）（a）和（b）的组合。更具体地说，本发明涉及一种方法，其中促进形成二硫化物新鲜的所述条件选自以下一种或多种：a）加入氧化剂，优选氧化金属离子，优选选自Cu2 + ，Fe2 +，Fe3 +和Zn2 +; b）结构域抗体与硫醇异构酶的共表达; c）使培养条件适应以下选自以下的一种或多种：降低培养温度和/或优化培养基，包括但不限于还原需要甲醇进料的宿主的甲醇进料，降低培养基的导电性，加入的酵母提取物和/或蛋白胨，或其任何组合; d）在变性剂和氧化还原缓冲剂存在下重折叠结构域抗体e）通过氧合，升高温度，增加pH，高压或其任何组合来处理结构域抗体，以及f）a）至e），或者其中除去缺少至少一个二硫键的结构域抗体的条件选自a）包含游离硫醇基团的结合域抗体适合于适当的反应性基团，包括但不限于固定硫醇基团，任选地在变性条件下; b）反相高效液相色谱法。

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