Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)

Autores
Paillet, Cristian; Forno, Guillermina; Soldano, Nicolas; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines. We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a threefold increase in trypsin concentration. The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 22 full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels. On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID50%/cell and trypsin concentration of 5 g/1.0 × 106 cells with a desirability of 0.737 was selected as the optimized condition for H1N1 production in sVero cells. Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.
Fil: Paillet, Cristian. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina
Fil: Forno, Guillermina. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Soldano, Nicolas. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; Argentina
Fil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
Influenza Virus
Vaccines
Vero Cells
Suspension Culture
Optimization
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/14440
Acceder
id CONICETDig_1dc04e53dcf81aa5dc5dcfbf2ffc87a1
oai_identifier_str oai:ri.conicet.gov.ar:11336/14440
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)Paillet, CristianForno, GuillerminaSoldano, NicolasKratje, Ricardo BertoldoEtcheverrigaray, MarinaInfluenza VirusVaccinesVero CellsSuspension CultureOptimizationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines. We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a threefold increase in trypsin concentration. The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 22 full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels. On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID50%/cell and trypsin concentration of 5 g/1.0 × 106 cells with a desirability of 0.737 was selected as the optimized condition for H1N1 production in sVero cells. Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.Fil: Paillet, Cristian. Zelltek S.A. Parque Tecnológico Litoral Centro; ArgentinaFil: Forno, Guillermina. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Soldano, Nicolas. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; ArgentinaFil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2011-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/14440Paillet, Cristian; Forno, Guillermina; Soldano, Nicolas; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero); Elsevier; Vaccine; 29; 41; 7-2011; 7212-72170264-410Xenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0264410X1info:eu-repo/semantics/altIdentifier/doi/10.1016/j.vaccine.2011.07.016info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:20:27Zoai:ri.conicet.gov.ar:11336/14440instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:20:27.717CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
title Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
spellingShingle Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
Paillet, Cristian
Influenza Virus
Vaccines
Vero Cells
Suspension Culture
Optimization
title_short Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
title_full Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
title_fullStr Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
title_full_unstemmed Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
title_sort Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
dc.creator.none.fl_str_mv Paillet, Cristian
Forno, Guillermina
Soldano, Nicolas
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
author Paillet, Cristian
author_facet Paillet, Cristian
Forno, Guillermina
Soldano, Nicolas
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
author_role author
author2 Forno, Guillermina
Soldano, Nicolas
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
author2_role author
author
author
author
dc.subject.none.fl_str_mv Influenza Virus
Vaccines
Vero Cells
Suspension Culture
Optimization
topic Influenza Virus
Vaccines
Vero Cells
Suspension Culture
Optimization
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines. We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a threefold increase in trypsin concentration. The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 22 full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels. On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID50%/cell and trypsin concentration of 5 g/1.0 × 106 cells with a desirability of 0.737 was selected as the optimized condition for H1N1 production in sVero cells. Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.
Fil: Paillet, Cristian. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina
Fil: Forno, Guillermina. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Soldano, Nicolas. Zelltek S.A. Parque Tecnológico Litoral Centro; Argentina; Argentina
Fil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines. We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a threefold increase in trypsin concentration. The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 22 full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels. On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID50%/cell and trypsin concentration of 5 g/1.0 × 106 cells with a desirability of 0.737 was selected as the optimized condition for H1N1 production in sVero cells. Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.
publishDate 2011
dc.date.none.fl_str_mv 2011-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/14440
Paillet, Cristian; Forno, Guillermina; Soldano, Nicolas; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero); Elsevier; Vaccine; 29; 41; 7-2011; 7212-7217
0264-410X
url http://hdl.handle.net/11336/14440
identifier_str_mv Paillet, Cristian; Forno, Guillermina; Soldano, Nicolas; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero); Elsevier; Vaccine; 29; 41; 7-2011; 7212-7217
0264-410X
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0264410X1
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.vaccine.2011.07.016
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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