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
- Institución
- Instituto Nacional de Tecnología Agropecuaria
- OAI Identificador
- oai:localhost:20.500.12123/4310
Ver los metadatos del registro completo
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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 |
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12.559606 |