Comparison of strategies for the production of FMDV empty capsids using the Baculovirus Vector System
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: infe...
| Autores principales: | , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2019
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| Materias: | |
| Acceso en línea: | 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 |
| _version_ | 1855483407958540288 |
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| author | Ruiz, Vanesa Mignaqui, Ana Clara Nuñez, M.C. Reytor, Edel Escribano, José M. Wigdorovitz, Andres |
| author_browse | Escribano, José M. Mignaqui, Ana Clara Nuñez, M.C. Reytor, Edel Ruiz, Vanesa Wigdorovitz, Andres |
| author_facet | Ruiz, Vanesa Mignaqui, Ana Clara Nuñez, M.C. Reytor, Edel Escribano, José M. Wigdorovitz, Andres |
| author_sort | Ruiz, Vanesa |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA4310 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA43102019-01-22T14:22:06Z Comparison of strategies for the production of FMDV empty capsids using the Baculovirus Vector System Ruiz, Vanesa Mignaqui, Ana Clara Nuñez, M.C. Reytor, Edel Escribano, José M. Wigdorovitz, Andres Enfermedades de los Animales Fiebre Aftosa Virus Fiebre Aftosa Baculovirus Células Animal Diseases Foot and Mouth Disease Aphthovirus Cells 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 2019-01-22T14:20:29Z 2019-01-22T14:20:29Z 2014-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng info:eu-repo/semantics/restrictedAccess application/pdf Springer Molecular Biotechnology 56 (11) : 963–970 (November 2014) |
| spellingShingle | Enfermedades de los Animales Fiebre Aftosa Virus Fiebre Aftosa Baculovirus Células Animal Diseases Foot and Mouth Disease Aphthovirus Cells Ruiz, Vanesa Mignaqui, Ana Clara Nuñez, M.C. Reytor, Edel Escribano, José M. Wigdorovitz, Andres 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 |
| 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_short | 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 |
| topic | Enfermedades de los Animales Fiebre Aftosa Virus Fiebre Aftosa Baculovirus Células Animal Diseases Foot and Mouth Disease Aphthovirus Cells |
| 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 |
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