Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System

Autores
Ruiz, Vanesa; Mignaqui, Ana Clara; Nuñez, M. C.; Reytor, E; Escribano, José M.; Wigdorovitz, Andrés
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Recombinant FMDV empty capsids have been produced in insect cells and larvae using the baculovirus expression system, although protein yield and efficiency of capsid assembly have been highly variable. In this work, two strategies were compared for the expression of FMDV A/Arg/01 empty capsids: infection with a dual-promoter baculovirus vector coding for the capsid precursor (P12A) and the protease 3C under the control of the polyhedrin and p10 promoters, respectively (BacP12A-3C), or a single-promoter vector coding the P12A3C cassette (BacP12A3C). Expression levels and assembly into empty capsids were analyzed in insect cells and larvae. We observed that the use of the single-promoter vector allowed higher levels of expression both in insect cells and larvae. Recombinant capsid proteins produced by both vectors were recognized by monoclonal antibodies (mAbs) directed against conformational epitopes of FMDV A/Arg/01 and proved to self-assemble into empty capsids (75S) and pentamers (12S) when analyzed by sucrose gradient centrifugation.
Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Nuñez, M. C.. Universidad Politécnica de Madrid; España
Fil: Reytor, E. Universidad Politécnica de Madrid; España
Fil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria ; España
Fil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
Foot And Mouth Disease Virus
Empty Capsids
Baculovirus Expression System
Sf9 Cells
T.Ni Larva
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/35648
Acceder
id CONICETDig_eedba6ddcbffe4a47c5e077759b549cf
oai_identifier_str oai:ri.conicet.gov.ar:11336/35648
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector SystemRuiz, VanesaMignaqui, Ana ClaraNuñez, M. C.Reytor, EEscribano, José M.Wigdorovitz, AndrésFoot And Mouth Disease VirusEmpty CapsidsBaculovirus Expression SystemSf9 CellsT.Ni Larvahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Recombinant FMDV empty capsids have been produced in insect cells and larvae using the baculovirus expression system, although protein yield and efficiency of capsid assembly have been highly variable. In this work, two strategies were compared for the expression of FMDV A/Arg/01 empty capsids: infection with a dual-promoter baculovirus vector coding for the capsid precursor (P12A) and the protease 3C under the control of the polyhedrin and p10 promoters, respectively (BacP12A-3C), or a single-promoter vector coding the P12A3C cassette (BacP12A3C). Expression levels and assembly into empty capsids were analyzed in insect cells and larvae. We observed that the use of the single-promoter vector allowed higher levels of expression both in insect cells and larvae. Recombinant capsid proteins produced by both vectors were recognized by monoclonal antibodies (mAbs) directed against conformational epitopes of FMDV A/Arg/01 and proved to self-assemble into empty capsids (75S) and pentamers (12S) when analyzed by sucrose gradient centrifugation.Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Nuñez, M. C.. Universidad Politécnica de Madrid; EspañaFil: Reytor, E. Universidad Politécnica de Madrid; EspañaFil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria ; EspañaFil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaSpringer2014-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/35648Ruiz, Vanesa; Mignaqui, Ana Clara; Nuñez, M. C.; Reytor, E; Escribano, José M.; et al.; Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System; Springer; Molecular Biotechnology; 56; 11; 6-2014; 963-9701073-60851559-0305CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1007/s12033-014-9775-8info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs12033-014-9775-8info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:39:55Zoai:ri.conicet.gov.ar:11336/35648instacron: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:34982025-09-29 10:39:55.682CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
title Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
spellingShingle Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
Ruiz, Vanesa
Foot And Mouth Disease Virus
Empty Capsids
Baculovirus Expression System
Sf9 Cells
T.Ni Larva
title_short Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
title_full Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
title_fullStr Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
title_full_unstemmed Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
title_sort Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System
dc.creator.none.fl_str_mv Ruiz, Vanesa
Mignaqui, Ana Clara
Nuñez, M. C.
Reytor, E
Escribano, José M.
Wigdorovitz, Andrés
author Ruiz, Vanesa
author_facet Ruiz, Vanesa
Mignaqui, Ana Clara
Nuñez, M. C.
Reytor, E
Escribano, José M.
Wigdorovitz, Andrés
author_role author
author2 Mignaqui, Ana Clara
Nuñez, M. C.
Reytor, E
Escribano, José M.
Wigdorovitz, Andrés
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Foot And Mouth Disease Virus
Empty Capsids
Baculovirus Expression System
Sf9 Cells
T.Ni Larva
topic Foot And Mouth Disease Virus
Empty Capsids
Baculovirus Expression System
Sf9 Cells
T.Ni Larva
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Recombinant FMDV empty capsids have been produced in insect cells and larvae using the baculovirus expression system, although protein yield and efficiency of capsid assembly have been highly variable. In this work, two strategies were compared for the expression of FMDV A/Arg/01 empty capsids: infection with a dual-promoter baculovirus vector coding for the capsid precursor (P12A) and the protease 3C under the control of the polyhedrin and p10 promoters, respectively (BacP12A-3C), or a single-promoter vector coding the P12A3C cassette (BacP12A3C). Expression levels and assembly into empty capsids were analyzed in insect cells and larvae. We observed that the use of the single-promoter vector allowed higher levels of expression both in insect cells and larvae. Recombinant capsid proteins produced by both vectors were recognized by monoclonal antibodies (mAbs) directed against conformational epitopes of FMDV A/Arg/01 and proved to self-assemble into empty capsids (75S) and pentamers (12S) when analyzed by sucrose gradient centrifugation.
Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Nuñez, M. C.. Universidad Politécnica de Madrid; España
Fil: Reytor, E. Universidad Politécnica de Madrid; España
Fil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria ; España
Fil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Recombinant FMDV empty capsids have been produced in insect cells and larvae using the baculovirus expression system, although protein yield and efficiency of capsid assembly have been highly variable. In this work, two strategies were compared for the expression of FMDV A/Arg/01 empty capsids: infection with a dual-promoter baculovirus vector coding for the capsid precursor (P12A) and the protease 3C under the control of the polyhedrin and p10 promoters, respectively (BacP12A-3C), or a single-promoter vector coding the P12A3C cassette (BacP12A3C). Expression levels and assembly into empty capsids were analyzed in insect cells and larvae. We observed that the use of the single-promoter vector allowed higher levels of expression both in insect cells and larvae. Recombinant capsid proteins produced by both vectors were recognized by monoclonal antibodies (mAbs) directed against conformational epitopes of FMDV A/Arg/01 and proved to self-assemble into empty capsids (75S) and pentamers (12S) when analyzed by sucrose gradient centrifugation.
publishDate 2014
dc.date.none.fl_str_mv 2014-06
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/11336/35648
Ruiz, Vanesa; Mignaqui, Ana Clara; Nuñez, M. C.; Reytor, E; Escribano, José M.; et al.; Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System; Springer; Molecular Biotechnology; 56; 11; 6-2014; 963-970
1073-6085
1559-0305
CONICET Digital
CONICET
url http://hdl.handle.net/11336/35648
identifier_str_mv Ruiz, Vanesa; Mignaqui, Ana Clara; Nuñez, M. C.; Reytor, E; Escribano, José M.; et al.; Comparison of Strategies for the Production of FMDV Empty Capsids Using the Baculovirus Vector System; Springer; Molecular Biotechnology; 56; 11; 6-2014; 963-970
1073-6085
1559-0305
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1007/s12033-014-9775-8
info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs12033-014-9775-8
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
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 13.070432

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