发明公开
EP1633860A2 IMPROVED METHOD FOR THE RECOVERY OF NON-SEGMENTED, NEGATIVE-STRANDED RNA VIRUSES FROM cDNA
审中-公开
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基本信息:
- 专利标题: IMPROVED METHOD FOR THE RECOVERY OF NON-SEGMENTED, NEGATIVE-STRANDED RNA VIRUSES FROM cDNA
- 专利标题(中):改进方法OBTAINING未分段负链RNA病毒从cDNA
- 申请号:EP04754803.7 申请日:2004-06-08
- 公开(公告)号:EP1633860A2 公开(公告)日:2006-03-15
- 发明人: PARKS, Christopher, L. , UDEM, Stephen, A. , SIDHU, Modinderjit, S.
- 申请人: Wyeth
- 申请人地址: 5 Giralda Farms Madison, NJ 07940-1027 US
- 专利权人: Wyeth
- 当前专利权人: Wyeth
- 当前专利权人地址: 5 Giralda Farms Madison, NJ 07940-1027 US
- 代理机构: Denholm, Anna Marie
- 优先权: US477389P 20030609
- 国际公布: WO2004113517 20041229
- 主分类号: C12N7/00
- IPC分类号: C12N7/00
摘要:
Methods for producing infectious, non-segmented, negative-stranded RNA viruses of the Order Mononegavirales are provided that involve coexpression of a viral cDNA along with essential viral proteins, N, P, and L in a host cell transiently transfected with an expression vector encoding an RNA polymerase. In alternate methods, after the host cell is transfected with a viral cDNA expression vector and one or more vectors encoding the RNA polymerase, N protein, P protein, and L protein, the host cell is exposed to an effective heat shock under conditions sufficient to increase recovery of the recombinant virus. In other alternate embodiments, the host cells are transferred after viral rescue begins into co-culture with a plaque expansion cell, typically a monolayer of expansion cells, and the assembled infectious, non-segmented, negative-stranded RNA virus is recovered from the co-culture. Also provided within the invention are compositions for producing infectious, non-segmented, negative-stranded RNA virus of the Order Mononegavirales, recombinant viruses produced using the foregoing methods and compositions, and immunogenic compositions and methods employing the recombinant viruses. In additional embodiments, the methods and compositions of the invention are employed to produce growth- or replication-defective non-segmented negative-stranded RNA viruses and subviral particles.