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61. 发明授权

US06265212B1 Packaging systems for human recombinant adenovirus to be used in gene therapy 有权
标题翻译：用于基因治疗的人重组腺病毒的包装系统
公开(公告)号：US06265212B1
公开(公告)日：2001-07-24
申请号：US09356575
申请日：1999-07-19
申请人： Frits J. Fallaux , Robert C. Hoeben , Abraham Bout , Domenico Valerio , Alex J. van der Eb , Govert Schouten
发明人： Frits J. Fallaux , Robert C. Hoeben , Abraham Bout , Domenico Valerio , Alex J. van der Eb , Govert Schouten
IPC分类号： C12N1563
CPC分类号： C12N7/00 , A61K48/00 , C07K14/005 , C12N15/86 , C12N2710/10322 , C12N2710/10343 , C12N2710/10352 , C12N2830/42
摘要： The problem of replication competent adenovirus in virus production is solved in that we have developed packaging cells that have no overlapping sequences with a new basic vector and, thus, are suited for safe large scale production of recombinant adenoviruses. One of the additional problems associated with the use of recombinant adenovirus vectors is the host-defense reaction against treatment with adenovirus. Another aspect of the invention involves screening recombinant adenovirus vector lots, especially those intended for clinical use, for the presence of adenovirus E1 sequences, as this will reveal replication competent adenovirus, as well as revertant E1 adenoviruses. It is also an aspect of the present invention to molecularly characterize the revertants that are generated in the newer helper/vector combinations.
摘要翻译：解决了病毒生产中复制能力的腺病毒的问题，因为我们已经开发了与新的碱性载体没有重叠序列的包装细胞，因此适合于重组腺病毒的安全大规模生产。与使用重组腺病毒载体相关的另外一个问题是用腺病毒治疗的宿主防御反应。本发明的另一方面涉及筛选腺病毒E1序列存在的重组腺病毒载体批次，特别是用于临床应用的重组腺病毒载体批次，因为这将揭示复制性腺病毒以及回复E1腺病毒。分子表征在较新的辅助/载体组合中产生的回复体也是本发明的一个方面。

62. 发明授权

US06033908A Packaging systems for human recombinant adenovirus to be used in gene therapy 失效
标题翻译：用于基因治疗的人重组腺病毒的包装系统
公开(公告)号：US06033908A
公开(公告)日：2000-03-07
申请号：US892873
申请日：1997-07-15
申请人： Abraham Bout , Robert Cornelis Hoeben
发明人： Abraham Bout , Robert Cornelis Hoeben
IPC分类号： C12N15/09 , A61K35/76 , A61K48/00 , C12N20060101 , C12N5/08 , C12N5/10 , C12N7/00 , C12N7/01 , C12N15/86 , C12N15/861 , C12N15/00
CPC分类号： C12N7/00 , C12N15/86 , A61K48/00 , C12N2710/10321 , C12N2710/10343 , C12N2710/10351 , C12N2710/10352 , C12N2830/00 , C12N2830/15 , C12N2830/42 , C12N2830/60
摘要： The invention provides improved methods and products based on adenoviral materials which can advantageously be used in for instance gene therapy. In one aspect an adenoviral vector is provided which has no overlap with a suitable packaging cell line which is another aspect of invention. This combination excludes the possibility of homologous recombination, thereby excluding the possibility of the formation of replication competent adenovirus. In another aspect an adenovirus based helper construct which by its size is incapable of being encapsidated. This helper virus can be transferred into any suitable host cell making it a packaging cell. Further a number of useful mutations to adenoviral based materials and combinations of such mutations are disclosed, which all have in common the safety of the methods and the products, in particular avoiding the production of replication competent adenovirus and/or interference with the immune system. Further a method of intracellular amplification is provided.
摘要翻译：本发明提供了可以有利地用于例如基因治疗的基于腺病毒材料的改进的方法和产品。在一个方面，提供了与适合的包装细胞系重叠的腺病毒载体，其是本发明的另一方面。该组合不包括同源重组的可能性，从而排除形成复制型腺病毒的可能性。在另一方面，基于腺病毒的辅助构建体，其大小不能被包被。这种辅助病毒可以转移到任何合适的宿主细胞中，使其成为包装细胞。此外，公开了一些对基于腺病毒的材料的有用的突变以及这些突变的组合，这些突变都具有方法和产物的安全性，特别是避免产生可复制的腺病毒和/或免疫系统的干扰。此外还提供了细胞内扩增的方法。

63. 发明授权

US5914231A Poly(organo)phosphazenes for use in synthetic transfection systems 失效
标题翻译：用于合成转染系统的聚（有机）磷腈
公开(公告)号：US5914231A
公开(公告)日：1999-06-22
申请号：US817912
申请日：1997-06-30
申请人： Wilhelmus Everhardus Hennink , Abraham Bout
发明人： Wilhelmus Everhardus Hennink , Abraham Bout
IPC分类号： C12N15/09 , A61K9/127 , A61K31/70 , A61K47/48 , A61K48/00 , C12N5/10 , C12N15/87 , C12N15/88 , C12Q1/68 , C07H21/04 , C12N15/63
CPC分类号： A61K47/48223 , C12N15/87 , A61K48/00
摘要： The present invention to a synthetic transfection system comprising as a carrier a cationic, water dispersible polyphosphazene. In addition, it relates to a method for introducing DNA fragments in target cells, comprising contacting these DNA fragments with a polyphosphazene which is at least partially substituted with cationic substituents and subsequently contacting the obtained transfection system with target cells. Finally, the invention involves the use of a polyphosphazene which is at least partially substituted with cationic substituents as a transfection vehicle.
摘要翻译： PCT No.PCT / NL96 / 00324 Sec。 371日期：1997年6月30日 102（e）日期1996年6月30日PCT 1996年8月16日PCT PCT。出版物WO97 / 07226 日期1997年2月27日本发明涉及一种合成转染系统，其包含阳离子，水分散性聚磷腈作为载体。此外，本发明涉及在靶细胞中引入DNA片段的方法，包括使这些DNA片段与至少部分被阳离子取代基部分取代的聚磷腈接触，随后使获得的转染系统与靶细胞接触。最后，本发明涉及使用至少部分被阳离子取代基取代的聚磷腈作为转染载体。

64. 发明授权

US08236561B2 Efficient production of IgA in recombinant mammalian cells 有权
标题翻译：在重组哺乳动物细胞中有效地产生IgA
公开(公告)号：US08236561B2
公开(公告)日：2012-08-07
申请号：US10644256
申请日：2003-08-20
申请人： David H. A. Jones , Abraham Bout
发明人： David H. A. Jones , Abraham Bout
IPC分类号： C12N5/071 , C12N15/85
CPC分类号： C07K14/005 , C12N15/86 , C12N2710/10322 , C12N2710/10343 , C12N2740/16234 , C12N2760/16122 , C12N2760/16134 , C12N2800/108 , C12N2830/00 , C12N2830/15 , C12N2830/60
摘要： The invention provides an immortalized human retina cell expressing E1A and E1B proteins of an adenovirus, wherein the cell has recombinant nucleic acid encoding an IgA molecule in expressible format. Also provided is a method for recombinant production of an IgA molecule, the method involving culturing a cell of the invention and expressing the recombinant nucleic acid encoding an IgA.
摘要翻译：本发明提供了表达腺病毒E1A和E1B蛋白的永生化人视网膜细胞，其中所述细胞具有编码可表达形式的IgA分子的重组核酸。还提供了重组生产IgA分子的方法，该方法涉及培养本发明的细胞并表达编码IgA的重组核酸。

65. 发明授权

US07968087B2 Gene delivery vectors provided with a tissue tropism for smooth muscle cells, and/or endothelial cells 有权
标题翻译：为平滑肌细胞和/或内皮细胞提供组织嗜性的基因递送载体
公开(公告)号：US07968087B2
公开(公告)日：2011-06-28
申请号：US12455086
申请日：2009-05-28
申请人： Ronald Vogels , Menzo J. E. Havenga , Abraham Bout
发明人： Ronald Vogels , Menzo J. E. Havenga , Abraham Bout
IPC分类号： A01N63/00 , A61K48/00 , C12N15/00 , C12N15/63 , C07H21/04
CPC分类号： C07K14/005 , A61K38/00 , A61K48/00 , C07K2319/00 , C12N15/86 , C12N2710/10322 , C12N2710/10343 , C12N2710/10345 , C12N2810/6018
摘要： A gene delivery vehicle having been provided with at least a tissue tropism for cells selected from the group of smooth muscle cells, endothelial cells, and/or liver cells. The tissue tropism is generally provided by a virus capsid, such as one comprising protein fragments from at least two different viruses, such as two different adenoviruses, including adenovirus of subgroup C or subgroup B (for example, adenovirus 16). The protein fragments can comprise a tissue tropism-determining fragment of a fiber protein derived from a subgroup B adenovirus. Also, cells for producing such gene delivery vehicles and pharmaceutical compositions containing these gene delivery vehicles are provided. Further, a method is disclosed for delivering nucleic acid to cells such as smooth muscle cells and/or endothelial cells which involves administering to the cells an adenovirus capsid having proteins from at least two different adenoviruses and wherein at least a tissue tropism-determining fragment of a fiber protein is derived from a subgroup B adenovirus. Particular constructs are also disclosed.
摘要翻译：具有至少一种选自平滑肌细胞，内皮细胞和/或肝细胞的细胞的组织嗜性的基因递送载体。组织向性通常由病毒衣壳提供，例如包含来自至少两种不同病毒的蛋白质片段的病毒衣壳，例如两种不同的腺病毒，包括亚组C或亚组B的腺病毒（例如，腺病毒16）。蛋白质片段可以包含衍生自亚组B腺病毒的纤维蛋白质的组织向性决定片段。此外，提供了用于产生这种基因递送载体的细胞和含有这些基因递送载体的药物组合物。此外，公开了一种用于将核酸递送至诸如平滑肌细胞和/或内皮细胞的细胞的方法，其涉及向细胞施用具有来自至少两种不同腺病毒的蛋白质的腺病毒衣壳，并且其中至少一种组织向性确定片段纤维蛋白源自B亚型腺病毒亚组。还公开了特定的构建体。

66. 发明授权

US07749493B2 Chimeric adenoviruses 失效
标题翻译：嵌合腺病毒
公开(公告)号：US07749493B2
公开(公告)日：2010-07-06
申请号：US11207626
申请日：2005-08-18
申请人： Menzo Havenga , Ronald Vogels , Abraham Bout
发明人： Menzo Havenga , Ronald Vogels , Abraham Bout
IPC分类号： A61K48/00 , C12N15/00 , C12N15/09 , C12N15/11 , C12N15/34
CPC分类号： C12N15/86 , C07K14/005 , C12N2710/10322 , C12N2710/10344 , C12N2760/20134
摘要： The present invention provides methods and vector systems for the generation of chimeric recombinant adenoviruses. These hybrid adenoviruses contain a genome that is derived from different adenovirus serotypes. In particular, novel hybrid adenoviruses are disclosed with improved properties for gene therapy purposes. These properties include: a decreased sensitivity towards neutralizing antibodies, a modified host range, a change in the titer to which adenovirus can be grown, the ability to escape trapping in the liver upon in vivo systemic delivery, and absence or decreased infection of antigen presenting cells (APC) of the immune system, such as macrophages or dendritic cells. These chimeric adenoviruses thus represent improved tools for gene therapy and vaccination since they overcome the limitations observed with the currently used serotype subgroup C adenoviruses.
摘要翻译：本发明提供用于产生嵌合重组腺病毒的方法和载体系统。这些杂合腺病毒含有衍生自不同腺病毒血清型的基因组。特别地，公开了具有改进的用于基因治疗目的的性质的新型杂合腺病毒。这些性质包括：对中和抗体的降低的敏感性，修饰的宿主范围，可以生长腺病毒的滴度的变化，在体内全身递送时逃避捕获在肝脏中的能力，以及抗原呈递的不存在或减少的感染免疫系统的细胞（APC），例如巨噬细胞或树突状细胞。因此，这些嵌合腺病毒代表了用于基因治疗和疫苗接种的改进工具，因为它们克服了目前使用的血清型亚型C腺病毒所观察到的限制。

67. 发明授权

US07696157B2 Methods and means for producing proteins with predetermined post-translational modifications 失效
标题翻译：用预定的翻译后修饰产生蛋白质的方法和方法
公开(公告)号：US07696157B2
公开(公告)日：2010-04-13
申请号：US11888776
申请日：2007-08-01
申请人： Dirk Jan Elbertus Opstelten , Johan Christiaan Kapteyn , Petrus Christianus Johannes Josephus Passier , Ronald Hendrik Peter Brus , Abraham Bout
发明人： Dirk Jan Elbertus Opstelten , Johan Christiaan Kapteyn , Petrus Christianus Johannes Josephus Passier , Ronald Hendrik Peter Brus , Abraham Bout
IPC分类号： A61K38/18 , C07K14/505
CPC分类号： C07K14/005 , A61K38/00 , C07K14/505 , C12N2710/10322 , C12N2710/10343 , C12P21/005
摘要： Described are methods for identifying, selecting, and obtaining mammalian cells capable of producing proteinaceous molecules having predetermined post-translational modifications, wherein the post-translational modifications are brought about by the mammalian cell in which the proteinaceous molecule is expressed. Preferably, the predetermined post-translational modifications include glycosylation. Also described are methods for obtaining and producing proteinaceous molecules, using mammalian cells obtainable by a method of the present invention. Preferably, the proteinaceous molecules include erythropoietin (EPO), since EPO's effect depends heavily on its glycosylation pattern. Mammalian cells that have been obtained on the basis of their ability to produce proteins and/or post-translational modifications that are indicative for a predetermined post-translational modification that is desired are also provided. Preferably, the mammalian cells have neural characteristics and properties, such that significant amounts of recombinant proteins can be produced that harbor “neural- or brain-type” properties.
摘要翻译：描述了用于鉴定，选择和获得能够产生具有预定的翻译后修饰的蛋白质分子的哺乳动物细胞的方法，其中翻译后修饰由其中表达蛋白质分子的哺乳动物细胞产生。优选地，预定的翻译后修饰包括糖基化。还描述了使用可通过本发明的方法获得的哺乳动物细胞获得和产生蛋白质分子的方法。优选地，蛋白质分子包括促红细胞生成素（EPO），因为EPO的作用很大程度上取决于其糖基化模式。还提供了基于其产生蛋白质的能力获得的哺乳动物细胞和/或指示期望的预定的翻译后修饰的翻译后修饰。优选地，哺乳动物细胞具有神经特征和性质，使得可以产生含有“神经或脑型”性质的显着量的重组蛋白。

68. 发明申请

US20090253207A1 Gene delivery vectors provided with a tissue tropism for smooth muscle cells, and/or endothelial cells 有权
标题翻译：为平滑肌细胞和/或内皮细胞提供组织嗜性的基因递送载体
公开(公告)号：US20090253207A1
公开(公告)日：2009-10-08
申请号：US12455086
申请日：2009-05-28
申请人： Ronald Vogels , Menzo J. E. Havenga , Abraham Bout
发明人： Ronald Vogels , Menzo J. E. Havenga , Abraham Bout
IPC分类号： C12N15/86
CPC分类号： C07K14/005 , A61K38/00 , A61K48/00 , C07K2319/00 , C12N15/86 , C12N2710/10322 , C12N2710/10343 , C12N2710/10345 , C12N2810/6018
摘要： A gene delivery vehicle having been provided with at least a tissue tropism for cells selected from the group of smooth muscle cells, endothelial cells, and/or liver cells. The tissue tropism is generally provided by a virus capsid, such as one comprising protein fragments from at least two different viruses, such as two different adenoviruses, including adenovirus of subgroup C or subgroup B (for example, adenovirus 16). The protein fragments can comprise a tissue tropism-determining fragment of a fiber protein derived from a subgroup B adenovirus. Also, cells for producing such gene delivery vehicles and pharmaceutical compositions containing these gene delivery vehicles are provided. Further, a method is disclosed for delivering nucleic acid to cells such as smooth muscle cells and/or endothelial cells which involves administering to the cells an adenovirus capsid having proteins from at least two different adenoviruses and wherein at least a tissue tropism-determining fragment of a fiber protein is derived from a subgroup B adenovirus. Particular constructs are also disclosed.
摘要翻译：具有至少一种选自平滑肌细胞，内皮细胞和/或肝细胞的细胞的组织嗜性的基因递送载体。组织向性通常由病毒衣壳提供，例如包含来自至少两种不同病毒的蛋白质片段的病毒衣壳，例如两种不同的腺病毒，包括亚组C或亚组B的腺病毒（例如，腺病毒16）。蛋白质片段可以包含衍生自亚组B腺病毒的纤维蛋白质的组织向性决定片段。此外，提供了用于产生这种基因递送载体的细胞和含有这些基因递送载体的药物组合物。此外，公开了一种用于将核酸递送至诸如平滑肌细胞和/或内皮细胞的细胞的方法，其涉及向细胞施用具有来自至少两种不同腺病毒的蛋白质的腺病毒衣壳，并且其中至少一种组织向性确定片段纤维蛋白源自B亚型腺病毒亚组。还公开了特定的构建体。

69. 发明授权

US07491532B2 Recombinant protein production in a human cell 有权
标题翻译：在人细胞中重组蛋白质的产生
公开(公告)号：US07491532B2
公开(公告)日：2009-02-17
申请号：US10790562
申请日：2004-03-01
申请人： Abraham Bout , Guus Hateboer , Karina Cornelia Verhulst , Alphonsus Gerardus Uytdehaag , Govert Johan Schouten
发明人： Abraham Bout , Guus Hateboer , Karina Cornelia Verhulst , Alphonsus Gerardus Uytdehaag , Govert Johan Schouten
IPC分类号： C12N15/85 , C12N15/00 , C12N15/09 , C12N15/63
CPC分类号： C12N15/86 , C07K14/005 , C12N2710/10322 , C12N2710/10332 , C12N2710/10343 , C12N2740/16122 , C12N2740/16234 , C12N2760/16122 , C12N2760/16134 , C12N2760/16151 , C12N2800/108 , C12N2830/00 , C12N2830/15 , C12N2830/60
摘要： Methods and compositions for the production of recombinant proteins in a eukaryotic cell line are disclosed. The methods and compositions are particularly useful for generating stable expression of recombinant proteins of interest that are modified post-translationally, for example, by glycosylation. Such proteins may have advantageous properties in comparison with their counterparts produced in non-human systems such as Chinese hamster ovary cells.
摘要翻译：公开了用于在真核细胞系中生产重组蛋白的方法和组合物。所述方法和组合物特别可用于产生翻译后修饰的重组蛋白质的稳定表达，例如通过糖基化。与在非人系统如中国仓鼠卵巢细胞中产生的对应物相比，这样的蛋白质可能具有有利的性质。

70. 发明申请

US20090023196A1 Stocks of replication-deficient adenovirus 审中-公开
标题翻译：复制缺陷腺病毒的库存
公开(公告)号：US20090023196A1
公开(公告)日：2009-01-22
申请号：US11879421
申请日：2007-07-16
申请人： Frits J. Fallaux , Robert C. Hoeben , Alex Jan van der Eb , Abraham Bout , Domenico Valerio
发明人： Frits J. Fallaux , Robert C. Hoeben , Alex Jan van der Eb , Abraham Bout , Domenico Valerio
IPC分类号： C12N7/01
CPC分类号： C12N7/00 , A61K48/00 , C12N15/86 , C12N2710/10321 , C12N2710/10343 , C12N2710/10351 , C12N2710/10352 , C12N2830/00 , C12N2830/15 , C12N2830/42 , C12N2830/60
摘要： Presented are ways to address the problem of replication-competent adenovirus in adenoviral production for use with, for example, gene therapy. Packaging cells having no overlapping sequences with a selected vector are suited for large scale production of recombinant adenoviruses. A system for use with the invention produces replication-defective adenovirus. The system includes a primary cell containing a nucleic acid based on or derived from adenovirus and an isolated recombinant nucleic acid molecule for transfer into the primary cell. The isolated recombinant nucleic acid molecule is based on or derived from an adenovirus, has at least one functional encapsidation signal and at least one functional Inverted Terminal Repeat, and lacks overlapping sequences with the nucleic acid of the cell. Otherwise, the overlapping sequences would enable homologous recombination leading to replication-competent adenovirus in the primary cell into which the isolated recombinant nucleic acid molecule is to be transferred.
摘要翻译：提出的方法是解决在例如基因治疗中使用的腺病毒生产中复制能力的腺病毒的问题。与所选载体不具有重叠序列的包装细胞适合大规模生产重组腺病毒。本发明使用的系统产生复制缺陷型腺病毒。该系统包括含有基于或衍生自腺病毒的核酸的原代细胞和用于转移至原代细胞的分离的重组核酸分子。分离的重组核酸分子基于或衍生自腺病毒，具有至少一个功能性包封信号和至少一个功能性反向终止重复，并且与细胞的核酸缺少重叠序列。否则，重叠序列将实现同源重组，导致分离的重组核酸分子将转移到其中的原代细胞中的复制能力的腺病毒。

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