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1. 发明申请

US20120058538A1 Helper-free Rescue of Recombinant Negative Strand RNA Virus Systems 审中-公开
标题翻译：无重力拯救重组负链RNA病毒系统
公开(公告)号：US20120058538A1
公开(公告)日：2012-03-08
申请号：US13270332
申请日：2011-10-11
申请人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
发明人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
IPC分类号： C12N7/00
CPC分类号： C12N7/00 , A61K39/00 , A61K48/00 , A61K2039/523 , A61K2039/525 , A61K2039/5256 , C07K14/005 , C12N15/86 , C12N2760/16143 , C12N2760/18122 , C12N2760/18143 , C12N2760/18151 , C12N2760/18152 , C12N2840/44 , Y02A50/466
摘要： The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and/or which express, package and/or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.
摘要翻译：本发明涉及通过完全基于载体的系统在辅助病毒辅助下在宿主细胞中产生感染性负链病毒的方法。特别地，本发明涉及在包含编码形成核糖核蛋白复合物（RNP）所必需的病毒蛋白的cDNA的表达载体和包含用于基因组病毒RNA的cDNA的表达载体的表达载体中，在不存在辅助病毒的情况下细胞内产生感染性重组负链RNA病毒的方法（vRNA）或相应的cRNA。本发明还涉及产生具有病毒基因突变和/或表达，包装和/或呈递由异源核酸序列编码的肽或多肽的感染性重链负链RNA病毒的方法。本发明还涉及本发明的重组负链RNA病毒或嵌合式负链RNA病毒在疫苗制剂和药物组合物中的用途。

2. 发明申请

US20090061521A1 Recombinant negative strand RNA virus expression systems and vaccines 审中-公开
标题翻译：重组负链RNA病毒表达系统和疫苗
公开(公告)号：US20090061521A1
公开(公告)日：2009-03-05
申请号：US12148711
申请日：2008-04-22
申请人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
发明人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
IPC分类号： C12N15/87 , C12N7/00
CPC分类号： C12N7/00 , A61K39/00 , A61K48/00 , A61K2039/523 , A61K2039/525 , A61K2039/5256 , C07K14/005 , C12N15/86 , C12N2760/16143 , C12N2760/18122 , C12N2760/18143 , C12N2760/18151 , C12N2760/18152 , C12N2840/44 , Y02A50/466
摘要： The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and/or which express, package and/or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.
摘要翻译：本发明涉及通过完全基于载体的系统在辅助病毒辅助下在宿主细胞中产生感染性负链病毒的方法。特别地，本发明涉及在包含编码形成核糖核蛋白复合物（RNP）所必需的病毒蛋白的cDNA的表达载体和包含用于基因组病毒RNA的cDNA的表达载体的表达载体中，在不存在辅助病毒的情况下细胞内产生感染性重组负链RNA病毒的方法（vRNA）或相应的cRNA。本发明还涉及产生具有病毒基因突变和/或表达，包装和/或呈递由异源核酸序列编码的肽或多肽的感染性重链负链RNA病毒的方法。本发明还涉及本发明的重组负链RNA病毒或嵌合式负链RNA病毒在疫苗制剂和药物组合物中的用途。

3. 发明授权

US06649372B1 Helper-free rescue of recombinant negative strand RNA virus 有权
标题翻译：无需辅助重组负链RNA病毒
公开(公告)号：US06649372B1
公开(公告)日：2003-11-18
申请号：US09724412
申请日：2000-11-28
申请人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
发明人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee , Ervin Fodor
IPC分类号： C12P2106
CPC分类号： C12N7/00 , A61K39/00 , A61K48/00 , A61K2039/523 , A61K2039/525 , A61K2039/5256 , C07K14/005 , C12N15/86 , C12N2760/16143 , C12N2760/18122 , C12N2760/18143 , C12N2760/18151 , C12N2760/18152 , C12N2840/44 , Y02A50/466
摘要： The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and/or which express, package and/or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.
摘要翻译：本发明涉及通过完全基于载体的系统在辅助病毒辅助下在宿主细胞中产生感染性负链病毒的方法。特别地，本发明涉及在包含编码形成核糖核蛋白复合物（RNP）所必需的病毒蛋白的cDNA的表达载体和包含用于基因组病毒RNA的cDNA的表达载体的表达载体中，在不存在辅助病毒的情况下细胞内产生感染性重组负链RNA病毒的方法（vRNA）或相应的cRNA。本发明还涉及产生具有病毒基因突变和/或表达，包装和/或呈递由异源核酸序列编码的肽或多肽的感染性重链负链RNA病毒的方法。本发明还涉及本发明的重组负链RNA病毒或嵌合式负链RNA病毒在疫苗制剂和药物组合物中的用途。

4. 发明授权

US06544785B1 Helper-free rescue of recombinant negative strand RNA viruses 有权
标题翻译：无需辅助重组负链RNA病毒
公开(公告)号：US06544785B1
公开(公告)日：2003-04-08
申请号：US09616527
申请日：2000-07-14
申请人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee
发明人： Peter Palese , Adolfo Garcia-Sastre , George G. Brownlee
IPC分类号： C12N500
CPC分类号： C12N7/00 , A61K39/00 , A61K48/00 , A61K2039/523 , A61K2039/525 , A61K2039/5256 , C07K14/005 , C12N15/86 , C12N2760/16143 , C12N2760/18122 , C12N2760/18143 , C12N2760/18151 , C12N2760/18152 , C12N2840/44 , Y02A50/466
摘要： The present invention relates methods of generating infectious negative-strand virus in host cells by an entirely vector-based system without the aid of a helper virus. In particular, the present invention relates methods of generating infectious recombinant negative-strand RNA viruses intracellularly in the absence of helper virus from expression vectors comprising cDNAs encoding the viral proteins necessary to form ribonucleoprotein complexes (RNPs) and expression vectors comprising cDNA for genomic viral RNA(s) (vRNAs) or the corresponding cRNA(s). The present invention also relates to methods of generating infectious recombinant negative-strand RNA viruses which have mutations in viral genes and/or which express, package and/or present peptides or polypeptides encoded by heterologous nucleic acid sequences. The present invention further relates the use of the recombinant negative-strand RNA viruses or chimeric negative-strand RNA viruses of the invention in vaccine formulations and pharmaceutical compositions.
摘要翻译：本发明涉及通过完全基于载体的系统在辅助病毒辅助下在宿主细胞中产生感染性负链病毒的方法。特别地，本发明涉及在包含编码形成核糖核蛋白复合物（RNP）所必需的病毒蛋白的cDNA的表达载体和包含用于基因组病毒RNA的cDNA的表达载体的表达载体中，在不存在辅助病毒的情况下细胞内产生感染性重组负链RNA病毒的方法（vRNA）或相应的cRNA。本发明还涉及产生具有病毒基因突变和/或表达，包装和/或呈递由异源核酸序列编码的肽或多肽的感染性重链负链RNA病毒的方法。本发明还涉及本发明的重组负链RNA病毒或嵌合式负链RNA病毒在疫苗制剂和药物组合物中的用途。

5. 发明授权

US5171569A Factor IX preparations uncontaminated by plasma components or pox virus 失效
标题翻译：因子IX制剂未被血浆成分或痘病毒污染
公开(公告)号：US5171569A
公开(公告)日：1992-12-15
申请号：US764073
申请日：1991-09-23
申请人： Donald S. Anson , George G. Brownlee , Ian M. Jones
发明人： Donald S. Anson , George G. Brownlee , Ian M. Jones
IPC分类号： A61K38/00 , C07K16/40 , C12N9/64
CPC分类号： C12N9/644 , C07K16/40 , C12Y304/21022 , A61K38/00
摘要： The blood-clotting protein, factor IX, is synthesized in the bod in liver cells, where it undergoes three distinct types of post-translational modification before it is secreted into the bloodstream as a 415 amino acid long protein. It is therefore a difficult protein to produce by recombinant DNA technology in a highly biologically active form. Nevertheless, such a result has been achieved by the present invention in which typically factor IX cDNA in a plasmid is linearized and inserted into an expression vector having a promoter sequence of SV40 early gene, an SV40 polyadenylation sequence, the TK/NEO selectable marker and an ampicillin resistance gene. Mammalian cells such as from a dog kidney or rat liver are transfected by the calcium phosphate precipitation method. High levels of factor IX in a fully or near-fully biologically active form, useful as a plasma-free preparation for treatment of patients suffering from Christmas Disease (haemophilia B), are obtainable without recourse to poxvirus vectors which would contaminate the protein.
摘要翻译：血液凝固蛋白，因子IX，在肝细胞中的骨骼中合成，其中它在作为415个氨基酸长的蛋白质分泌到血液中之前经历三种不同类型的翻译后修饰。因此，通过重组DNA技术以高度生物活性形式生产是困难的蛋白质。然而，通过本发明已经实现了这样的结果，其中通常将质粒中的因子IX cDNA线性化并插入具有SV40早期基因启动子序列，SV40多聚腺苷酸化序列，TK / NEO选择标记的表达载体中，氨苄青霉素抗性基因。哺乳动物细胞如狗肾或大鼠肝脏通过磷酸钙沉淀法转染。完全或接近完全生物活性形式的高水平因子IX可用作治疗患有圣诞节疾病（血友病B）的患者的无血浆制剂，无需求助于污染蛋白质的痘病毒载体。

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