Factors influencing the production of recombinant SV40 vectors

Most gene therapy approaches employ viral vectors for gene delivery. Ideally, these vectors should be produced at high titer and purity with well-established protocols. Standardized methods to measure the quality of the vectors produced are imperative, as are techniques that allow reproducible quant...

ver descrição completa

Detalhes bibliográficos
Autores: Vera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385, Prieto, J. (Jesús)|||/items/0d9c3dec-4a09-400d-8c83-23ece1096c71, Strayer, D.S. (David S.)|||/items/f1bc8310-06be-4359-8447-7723af40885e, Fortes, P. (Puri)|||/items/4b719a39-983d-402a-bb2a-2bdd276be18e
Formato: artículo
Fecha de publicación:2004
País:España
Recursos:Universidad de Navarra
Repositorio:Dadun. Depósito Académico Digital de la Universidad de Navarra
Idioma:inglés
OAI Identifier:oai:dadun.unav.edu:10171/23152
Acesso em linha:https://hdl.handle.net/10171/23152
Access Level:acceso abierto
Palavra-chave:SV40
Recombinant SV40 vectors
Gene therapy
Production
Titration
DI particles
wtSV40 revertants
id ES_9ccb73a84fce58ed1f53f8df6dcaa47e
oai_identifier_str oai:dadun.unav.edu:10171/23152
network_acronym_str ES
network_name_str España
repository_id_str
spelling Factors influencing the production of recombinant SV40 vectorsVera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385Prieto, J. (Jesús)|||/items/0d9c3dec-4a09-400d-8c83-23ece1096c71Strayer, D.S. (David S.)|||/items/f1bc8310-06be-4359-8447-7723af40885eFortes, P. (Puri)|||/items/4b719a39-983d-402a-bb2a-2bdd276be18eSV40Recombinant SV40 vectorsGene therapyProductionTitrationDI particleswtSV40 revertantsMost gene therapy approaches employ viral vectors for gene delivery. Ideally, these vectors should be produced at high titer and purity with well-established protocols. Standardized methods to measure the quality of the vectors produced are imperative, as are techniques that allow reproducible quantitation of viral titer. We devised a series of protocols that achieve high-titer production and reproducible purification and provide for quality control and titering of recombinant simian virus 40 vectors (rSV40s). rSV40s are good candidate vehicles for gene transfer: they are easily modified to be nonreplicative and they are nonimmunogenic. Further, they infect a wide variety of cells and allow long-term transgene expression. We report here these protocols to produce rSV40 vectors in high yields, describe their purification, and characterize viral stocks using quality control techniques that monitor the presence of wild-type SV40 revertants and defective interfering particles. Several methods for reproducible titration of rSV40 viruses have been compared. We believe that these techniques can be widely applied to obtain high concentrations of high-quality rSV40 viruses reproducibly.Nature Publishing GroupDadun. Depósito Académico Digital Universidad de Navarra20122012-09-2020042004-01-0120042004-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10171/23152reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/231522026-06-21T12:47:57Z
dc.title.none.fl_str_mv Factors influencing the production of recombinant SV40 vectors
title Factors influencing the production of recombinant SV40 vectors
spellingShingle Factors influencing the production of recombinant SV40 vectors
Vera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385
SV40
Recombinant SV40 vectors
Gene therapy
Production
Titration
DI particles
wtSV40 revertants
title_short Factors influencing the production of recombinant SV40 vectors
title_full Factors influencing the production of recombinant SV40 vectors
title_fullStr Factors influencing the production of recombinant SV40 vectors
title_full_unstemmed Factors influencing the production of recombinant SV40 vectors
title_sort Factors influencing the production of recombinant SV40 vectors
dc.creator.none.fl_str_mv Vera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385
Prieto, J. (Jesús)|||/items/0d9c3dec-4a09-400d-8c83-23ece1096c71
Strayer, D.S. (David S.)|||/items/f1bc8310-06be-4359-8447-7723af40885e
Fortes, P. (Puri)|||/items/4b719a39-983d-402a-bb2a-2bdd276be18e
author Vera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385
author_facet Vera, M. (María)|||/items/8f8ce09a-134e-4bcb-9c35-f04d2477d385
Prieto, J. (Jesús)|||/items/0d9c3dec-4a09-400d-8c83-23ece1096c71
Strayer, D.S. (David S.)|||/items/f1bc8310-06be-4359-8447-7723af40885e
Fortes, P. (Puri)|||/items/4b719a39-983d-402a-bb2a-2bdd276be18e
author_role author
author2 Prieto, J. (Jesús)|||/items/0d9c3dec-4a09-400d-8c83-23ece1096c71
Strayer, D.S. (David S.)|||/items/f1bc8310-06be-4359-8447-7723af40885e
Fortes, P. (Puri)|||/items/4b719a39-983d-402a-bb2a-2bdd276be18e
author2_role author
author
author
dc.contributor.none.fl_str_mv Dadun. Depósito Académico Digital Universidad de Navarra
dc.subject.none.fl_str_mv SV40
Recombinant SV40 vectors
Gene therapy
Production
Titration
DI particles
wtSV40 revertants
topic SV40
Recombinant SV40 vectors
Gene therapy
Production
Titration
DI particles
wtSV40 revertants
description Most gene therapy approaches employ viral vectors for gene delivery. Ideally, these vectors should be produced at high titer and purity with well-established protocols. Standardized methods to measure the quality of the vectors produced are imperative, as are techniques that allow reproducible quantitation of viral titer. We devised a series of protocols that achieve high-titer production and reproducible purification and provide for quality control and titering of recombinant simian virus 40 vectors (rSV40s). rSV40s are good candidate vehicles for gene transfer: they are easily modified to be nonreplicative and they are nonimmunogenic. Further, they infect a wide variety of cells and allow long-term transgene expression. We report here these protocols to produce rSV40 vectors in high yields, describe their purification, and characterize viral stocks using quality control techniques that monitor the presence of wild-type SV40 revertants and defective interfering particles. Several methods for reproducible titration of rSV40 viruses have been compared. We believe that these techniques can be widely applied to obtain high concentrations of high-quality rSV40 viruses reproducibly.
publishDate 2004
dc.date.none.fl_str_mv 2004
2004-01-01
2004
2004-01-01
2012
2012-09-20
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/10171/23152
url https://hdl.handle.net/10171/23152
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra
instname:Universidad de Navarra
instname_str Universidad de Navarra
reponame_str Dadun. Depósito Académico Digital de la Universidad de Navarra
collection Dadun. Depósito Académico Digital de la Universidad de Navarra
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869414688900513792
score 15,300719

Registos relacionados