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, Edel; Escribano, José M.; Wigdorovitz, Andres
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.
Instituto de Virología
Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Nuñez, M.C. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); España
Fil: Reytor, Edel. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); España
Fil: Escribano, José M. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). Departamento de Biotecnología; España
Fil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Molecular Biotechnology 56 (11) : 963–970 (November 2014)
Materia
Enfermedades de los Animales
Fiebre Aftosa
Virus Fiebre Aftosa
Baculovirus
Células
Animal Diseases
Foot and Mouth Disease
Aphthovirus
Cells
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/4310

id INTADig_f296dfb46a08ad7b981e058221f0e023
oai_identifier_str oai:localhost:20.500.12123/4310
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Comparison of strategies for the production of FMDV empty capsids using the Baculovirus Vector SystemRuiz, VanesaMignaqui, Ana ClaraNuñez, M.C.Reytor, EdelEscribano, José M.Wigdorovitz, AndresEnfermedades de los AnimalesFiebre AftosaVirus Fiebre AftosaBaculovirusCélulasAnimal DiseasesFoot and Mouth DiseaseAphthovirusCellsRecombinant 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.Instituto de VirologíaFil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Nuñez, M.C. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); EspañaFil: Reytor, Edel. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); EspañaFil: Escribano, José M. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). Departamento de Biotecnología; EspañaFil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaSpringer2019-01-22T14:20:29Z2019-01-22T14:20:29Z2014-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://link.springer.com/article/10.1007/s12033-014-9775-8http://hdl.handle.net/20.500.12123/43101073-60851559-0305https://doi.org/10.1007/s12033-014-9775-8Molecular Biotechnology 56 (11) : 963–970 (November 2014)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-09-29T13:44:33Zoai:localhost:20.500.12123/4310instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:44:34.051INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
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
Enfermedades de los Animales
Fiebre Aftosa
Virus Fiebre Aftosa
Baculovirus
Células
Animal Diseases
Foot and Mouth Disease
Aphthovirus
Cells
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, Edel
Escribano, José M.
Wigdorovitz, Andres
author Ruiz, Vanesa
author_facet Ruiz, Vanesa
Mignaqui, Ana Clara
Nuñez, M.C.
Reytor, Edel
Escribano, José M.
Wigdorovitz, Andres
author_role author
author2 Mignaqui, Ana Clara
Nuñez, M.C.
Reytor, Edel
Escribano, José M.
Wigdorovitz, Andres
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Enfermedades de los Animales
Fiebre Aftosa
Virus Fiebre Aftosa
Baculovirus
Células
Animal Diseases
Foot and Mouth Disease
Aphthovirus
Cells
topic Enfermedades de los Animales
Fiebre Aftosa
Virus Fiebre Aftosa
Baculovirus
Células
Animal Diseases
Foot and Mouth Disease
Aphthovirus
Cells
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.
Instituto de Virología
Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Nuñez, M.C. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); España
Fil: Reytor, Edel. Universidad Politécnica de Madrid. Parque Científico y Tecnológico. Alternative Gene Expression S.L. (ALGENEX); España
Fil: Escribano, José M. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). Departamento de Biotecnología; España
Fil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). 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 2014-11
2019-01-22T14:20:29Z
2019-01-22T14:20:29Z
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 https://link.springer.com/article/10.1007/s12033-014-9775-8
http://hdl.handle.net/20.500.12123/4310
1073-6085
1559-0305
https://doi.org/10.1007/s12033-014-9775-8
url https://link.springer.com/article/10.1007/s12033-014-9775-8
http://hdl.handle.net/20.500.12123/4310
https://doi.org/10.1007/s12033-014-9775-8
identifier_str_mv 1073-6085
1559-0305
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Molecular Biotechnology 56 (11) : 963–970 (November 2014)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
_version_ 1844619130118864896
score 12.559606