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公开(公告)号：EP1507863B1
公开(公告)日：2006-12-27
申请号：EP03752862.7
申请日：2003-05-19
发明人： HANAK, Julian, Cobra Therapeutics Limited , CRANENBURGH, Rocky, Cobra Therapeutics Limited , GORMAN, Scott, Pharmaventures Limited
IPC分类号： C12N15/68 , C12N15/74 , C12N15/64 , A61K39/112 , A61K39/02 , A61K35/74 , A61K48/00 , C12N1/36 , C12R1/42
CPC分类号： C12N1/36 , A61K48/00 , A61K2035/11 , A61K2039/522 , C12N15/64 , C12N15/68 , C12N15/74 , Y02A50/484
摘要： This invention provides in vivo methods of maintaining a plasmid within a host cell, which is in turn within a recipient organism. In particular it relates to methods of maintaining a plasmid containing a gene useful in therapy, especially vaccination. The invention is based on modified host cells, suitable for in vivo therapeutic use, that may be used to express a therapeutically useful plasmid-borne gene, using a plasmid maintenance system that does not require the use of a plasmid-borne dominant selectable marker, but rather utilises a system of repressor titration. Methods of selecting host cells comprising the plasmid and host cells for use in these methods are also provided.
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公开(公告)号：EP1766036A1
公开(公告)日：2007-03-28
申请号：EP05762102.1
申请日：2005-07-01
发明人： CRANENBURGH, Rocky Marc Cobra Biologics Limited , BLOOR, Alexandra Elizabeth Cobra Biologics Limited
IPC分类号： C12N15/90
CPC分类号： C12N15/86 , C12N2740/13043 , C12N2740/13045 , C12N2800/30
摘要： The invention relates to a process for the removal of selectable marker gene sequences, in particular antibiotic gene sequences, from nucleic acid molecules, wherein the process makes use of dif-like site-specific recombinase recognition sites. The invention further relates to the application of this process in the unlabelled integration and deletion of chromosomal genes and in controlling gene expression.
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公开(公告)号：EP1697522A2
公开(公告)日：2006-09-06
申请号：EP04798638.5
申请日：2004-11-22
CPC分类号： C12N15/72 , A61K2035/11
摘要： The invention relates to a system for stable maintenance of a plasmid, to host cells for use in this system and to methods of using the system to obtain a plasmid useful in medical applications. In particular, the invention provides transformed host cell containing: i) a chromosomal gene which inhibits cell growth; and ii) a plasmid encoding an antisense sequence, wherein the antisense sequence encoded by the plasmid inhibits the action of the chromosomal gene, thereby permitting cell growth and a method for stable maintenance of a plasmid in a host cell in vivo.
摘要翻译： 本发明涉及稳定维持质粒的系统，用于该系统的宿主细胞以及使用该系统获得用于医学应用的质粒的方法。 特别地，本发明提供了转化的宿主细胞，其包含：i）抑制细胞生长的染色体基因; 和ii）编码反义序列的质粒，其中由质粒编码的反义序列抑制染色体基因的作用，由此允许细胞生长，以及在体内稳定维持宿主细胞中质粒的方法。
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公开(公告)号：EP0880585B1
公开(公告)日：2006-05-24
申请号：EP97903440.2
申请日：1997-02-12
发明人： THATCHER, David, Robert , HITCHCOCK, Antony, Gordon , HANAK, Julian, Alexis , VARLEY, Diane, Lesley
CPC分类号： C12N1/06 , C12N15/101 , C12N15/70
摘要： A scalable method for the production of highly purified plasmid DNA in Escherichia coli is described, which method includes growing plasmid-containing cells to a high biomass in exponential growth and lysing the cells by raising the pH of the culture to a carefully controlled pH value in which chromosomal DNA is denatured but plasmid DNA is reversibly renatured. The method has been developed for the production of pharmaceutical grade DNA for use in in vivo and ex vivo gene therapy.
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公开(公告)号：EP2588615A1
公开(公告)日：2013-05-08
申请号：EP11733896.2
申请日：2011-06-28
IPC分类号： C12N15/64
CPC分类号： C12N15/64
摘要： A method of producing a selectable marker gene-free plasmid by culturing a plasmid containing a selectable marker gene flanked by site specific recombinase target sites in a host cell environment incapable of effecting recombination between the site specific recombinase target sites and subsequently culturing the plasmid in another host cell environment which is capable of effecting recombination between the site specific recombinase target sites, so that the selectable marker gene is excised. Uses of plasmids produced by the method for the production of recombinant protein for therapeutic and vaccine purposes, production of therapeutic DNA and DNA vaccines and delivery of recombinant protein and DNA to a patient using live bacterial vectors.
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公开(公告)号：EP1697522B1
公开(公告)日：2010-07-14
申请号：EP04798638.5
申请日：2004-11-22
CPC分类号： C12N15/72 , A61K2035/11
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公开(公告)号：EP1670913A1
公开(公告)日：2006-06-21
申请号：EP04768843.7
申请日：2004-10-11
IPC分类号： C12N15/10
CPC分类号： C12N15/101
摘要： The invention relates to the purification of biological macromolecules from biological feedstocks by means of ion exchange chromatography. In particular, it relates to methods of purifying plasmid DNA from cell lysates, preferably without the need to pre-treat the cell lysate to remove RNA.
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9.发明授权
公开(公告)号：EP1080179B1
公开(公告)日：2006-06-07
申请号：EP99915927.0
申请日：1999-04-13
发明人： HANAK, Julian Alexis John, Cobra Therapeutics Ltd , WILLIAMS, Steven Geraint, Cobra Therapeutics Ltd
摘要： The invention relates to host cells and methods of preparing a substantially RNA-free cellular component, comprising culturing cells producing the cellular component in a medium and lysing the cells to produce a cell lysate, wherein the cell lysate contains the cellular component and sufficient RNase activity to degrade substantially all of the RNA molecules present in the cell lysate. The invention also relates to substantially RNA-free cellular components.
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公开(公告)号：EP1398072A1
公开(公告)日：2004-03-17
申请号：EP03078097.7
申请日：2000-03-10
CPC分类号： C12M47/06 , B01F7/1675 , B01F7/183 , B01F15/00201 , C12N1/066
摘要： The invention relates to a vessel for mixing a cell lysate. The invention also relates to a method of using the vessel to mix a cell lysate in order to obtain high-purity products such as nucleic acids or proteins for use in a variety of applications. The invention also relates to a method for monitoring the degree of cell lysis in a cell suspension by measuring the viscosity of the cell suspension.
摘要翻译： 一种用于混合含有染色体DNA的细胞悬浮液/裂解物的容器和包含一个或多个低功率数量叶轮的期望产物，其定位使得它们能够通过容器的内容物均匀地散发能量。 一种用于混合含有染色体DNA的细胞悬浮液/裂解物的容器和包含一个或多个低功率数量叶轮的期望产物，其定位使得它们能够通过容器的内容物均匀地散发能量。 容器还包括：（a）一个或多个进料管线，其能够将流体添加到围绕一个或多个叶轮的远端形成的良好混合区域; 和（b）围绕容器的内表面基本上均匀分布的一个或多个挡板，其中挡板基本上平行于叶轮的旋转轴线延伸。 独立权利要求还包括以下内容：（1）一种用于监测通过旋转一个或多个叶轮来混合其内容物的容器中的细胞悬浮液的裂解程度的方法，包括测量施加到叶轮轴上的扭矩，其中 当其稳定在最大值时，扭矩与细胞裂解程度成比例地增加直到裂解完成; （2）从新型容器中的细胞悬浮液制备所需产物的方法; （3）从使用新血管裂解的细胞培养物或（1）的方法得到的质粒或蛋白质; （4）包括螺旋叶轮的两个或多个部分的螺旋状叶轮的分段螺旋叶轮，当存在于轴上时，在不包含叶片或叶轮的叶片的一部分的部分之间具有间隙。
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