Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells
In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (GE) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chim...
| Autores principales: | , , , , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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Springer
2020
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/6676 https://link.springer.com/article/10.1007/s13205-019-1920-4 https://doi.org/10.1007/s13205-019-1920-4 |
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| author | Targovnik, Alexandra Marisa Ferrari, Alejandro Mc Callum, Gregorio Juan Arregui, Mariana Bernadett Smith, Ignacio Bracco, Lautaro Fidel Alfonso, Victoria Lopez, Maria Gabriela Martínez‑Solís, María Herrero, Salvador Miranda, Maria Victoria |
| author_browse | Alfonso, Victoria Arregui, Mariana Bernadett Bracco, Lautaro Fidel Ferrari, Alejandro Herrero, Salvador Lopez, Maria Gabriela Martínez‑Solís, María Mc Callum, Gregorio Juan Miranda, Maria Victoria Smith, Ignacio Targovnik, Alexandra Marisa |
| author_facet | Targovnik, Alexandra Marisa Ferrari, Alejandro Mc Callum, Gregorio Juan Arregui, Mariana Bernadett Smith, Ignacio Bracco, Lautaro Fidel Alfonso, Victoria Lopez, Maria Gabriela Martínez‑Solís, María Herrero, Salvador Miranda, Maria Victoria |
| author_sort | Targovnik, Alexandra Marisa |
| collection | INTA Digital |
| description | In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (GE) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (polh-pSeL), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard polh promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, GE was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein. |
| format | Artículo |
| id | INTA6676 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA66762020-01-14T15:31:24Z Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells Targovnik, Alexandra Marisa Ferrari, Alejandro Mc Callum, Gregorio Juan Arregui, Mariana Bernadett Smith, Ignacio Bracco, Lautaro Fidel Alfonso, Victoria Lopez, Maria Gabriela Martínez‑Solís, María Herrero, Salvador Miranda, Maria Victoria Baculovirus Virus de la Rabia Glicoproteínas Anticuerpos Rabies Virus Glycoproteins Antibodies In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (GE) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (polh-pSeL), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard polh promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, GE was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein. Instituto de Biotecnología Fil: Targovnik, Alexandra Marisa. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ferrari, Alejandro. Universidad de Buenos Aires. Instituto de Química y Físico-Química Biológicas “Prof. Alejandro C. Paladini”; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mc Callum, Gregorio Juan. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Arregui, Mariana Bernadett. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Smith, Ignacio. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Bracco, Lautaro Fidel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Martínez‑Solís, María. Universitat de València. Department of Genetics and Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina; España Fil: Herrero, Salvador. Universitat de València. Department of Genetics and Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina; España Fil: Miranda, Maria Victoria. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2020-01-14T15:24:49Z 2020-01-14T15:24:49Z 2019-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/6676 https://link.springer.com/article/10.1007/s13205-019-1920-4 2190-5738 https://doi.org/10.1007/s13205-019-1920-4 eng info:eu-repo/semantics/restrictedAccess application/pdf Springer 3 Biotech 9 : 385. (Noviembre 2019) |
| spellingShingle | Baculovirus Virus de la Rabia Glicoproteínas Anticuerpos Rabies Virus Glycoproteins Antibodies Targovnik, Alexandra Marisa Ferrari, Alejandro Mc Callum, Gregorio Juan Arregui, Mariana Bernadett Smith, Ignacio Bracco, Lautaro Fidel Alfonso, Victoria Lopez, Maria Gabriela Martínez‑Solís, María Herrero, Salvador Miranda, Maria Victoria Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title_full | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title_fullStr | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title_full_unstemmed | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title_short | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
| title_sort | highly efficient production of rabies virus glycoprotein g ectodomain in sf9 insect cells |
| topic | Baculovirus Virus de la Rabia Glicoproteínas Anticuerpos Rabies Virus Glycoproteins Antibodies |
| url | http://hdl.handle.net/20.500.12123/6676 https://link.springer.com/article/10.1007/s13205-019-1920-4 https://doi.org/10.1007/s13205-019-1920-4 |
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