公开(公告)号：EP1633860A2

公开(公告)日：2006-03-15

申请号：EP04754803.7

申请日：2004-06-08

申请人： Wyeth

发明人： PARKS, Christopher, L. ,  UDEM, Stephen, A. ,  SIDHU, Modinderjit, S.

IPC分类号： C12N7/00

CPC分类号： C07K14/005 ,  A61K39/00 ,  A61K48/00 ,  A61K2039/5254 ,  A61K2039/5256 ,  A61K2039/5258 ,  C12N7/00 ,  C12N15/86 ,  C12N2760/18022 ,  C12N2760/18422 ,  C12N2760/18451 ,  C12N2760/18452 ,  C12N2760/18551 ,  C12N2760/18552 ,  C12N2760/18622 ,  C12N2760/18651 ,  C12N2760/18652 ,  C12N2760/18662 ,  C12N2760/20243 ,  C12N2760/20251 ,  C12N2760/20252 ,  C12N2760/20262

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.

公开(公告)号：EP1292615B1

公开(公告)日：2006-10-25

申请号：EP01944623.6

申请日：2001-06-21

申请人： Wyeth Holdings Corporation

发明人： PARKS, Christopher, L.

IPC分类号： C07K14/115 ,  C12N7/00 ,  C07K14/13 ,  A61K39/165

CPC分类号： C07K14/005 ,  A61K39/00 ,  A61K39/12 ,  A61K39/165 ,  A61K2039/525 ,  A61K2039/5254 ,  C07K2319/00 ,  C12N7/00 ,  C12N2760/18422 ,  C12N2760/18434 ,  C12N2760/18461

摘要： Modified Morbilliviruses having at least one mutation in the region corresponding to amino acids 112-134 of the measles virus V protein are described wherein one or both of amino acids 113 or 114 is mutated. Such modified Morbilliviruses exhibit reduced repression of gene expression. Additional mutations or deletions in other regions of the genome may be included, including in the carboxy-terminal region.

公开(公告)号：EP1090108B1

公开(公告)日：2011-02-23

申请号：EP99927175.2

申请日：1999-06-03

申请人： Wyeth Holdings Corporation

发明人： PARKS, Christopher, L. ,  SIDHU, Mohinderjit, S. ,  UDEM, Stephen, A. ,  KOVACS, Gerald, R.

IPC分类号： C12N7/02 ,  C12N7/04 ,  C12N15/86 ,  C07K14/145

CPC分类号： C12N15/86 ,  C12N2710/24143 ,  C12N2830/002 ,  C12N2830/60 ,  C12N2830/85 ,  C12N2840/203

摘要： This invention relates to improved methods for producing nonsegmented, negative-sense, single-stranded RNA viruses of the Order designated Mononegavirales virus, including embodiments relating to methods of producing such viruses as attenuated and/or infectious viruses, such as Measles virus (MV) and respiratory syncytial virus (RSV). One method for producing a recombinant virus from the Order Mononegavirales comprises (a) in at least one host cell, conducting transfection of a rescue composition which comprises (i) a transcription vector comprising an isolated nucleic acid molecule which comprises polynucleotide sequence encoding a genome or antigenome of a nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales and (ii) at least one expression vector which comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins necessary for encapsidation, transcription and replication; in a host cell under conditions sufficient to permit the co-expression of these vectors and the production of the recombinant virus; (b) heating the transfected rescue composition to an effective heat shock temperature under conditions sufficient to increase the recovery of the recombinant virus; and optionally, (c) harvesting the resulting recombinant virus.

公开(公告)号：EP2225368A1

公开(公告)日：2010-09-08

申请号：EP08868608.4

申请日：2008-12-18

申请人： Wyeth LLC

发明人： PARKS, Christopher, L. ,  WITKO, Susan, E. ,  SIDHU, Maninder, K. ,  JOHNSON, J. Erik ,  HENDRY, Roger, Michael

IPC分类号： C12N7/04 ,  C07K14/145

CPC分类号： C12N7/00 ,  C07K14/005 ,  C12N2760/20222 ,  C12N2760/20252 ,  C12N2760/20261

摘要： A method of producing propagation-defective Vesicular Stomatitis Virus (VSV) in a cell culture is provided. The method involves introducing a plasmid vector encoding an optimized VSV G gene into a cell; expressing VSV G protein from the optimized VSV G gene; and introducing a propagation-defective VSV into the cell expressing the VSV G protein encoded by the optimized VSV G gene. The method further includes growing the cells in culture; and recovering the propagation-defective VSV from the culture.

公开(公告)号：EP2225264A1

公开(公告)日：2010-09-08

申请号：EP08867848.7

申请日：2008-12-18

申请人： Wyeth LLC

发明人： PARKS, Christopher, L. ,  WITKO, Susan, E. ,  SIDHU, Maninder, K. ,  JOHNSON, J., Erik ,  HENDRY, Roger, M.

IPC分类号： C07K14/145 ,  C12N7/04 ,  C12N15/63

CPC分类号： C12N7/00 ,  C12N2510/02 ,  C12N2760/20251 ,  C12N2760/20252

摘要： A method of producing propagation-defective Vesicular Stomatitis Virus (VSV) is provided. The method involves providing a cell that includes an optimized VSV G gene, wherein expression of VSV G protein from the optimized VSV G gene is inducible; and inducing the cell to express VSV G protein from the optimized VSV G gene. The method also involves infecting the induced cell with an attenuated VSV; growing the infected cells in culture; and recovering attenuated VSV from the culture.