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: | , , , , , , , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2019 |
| País: | Argentina |
| Institución: | Consejo Nacional de Investigaciones Científicas y Técnicas |
| Repositorio: | CONICET Digital (CONICET) |
| Idioma: | inglés |
| OAI Identifier: | oai:ri.conicet.gov.ar:11336/121583 |
| Acceso en línea: | http://hdl.handle.net/11336/121583 |
| Access Level: | acceso abierto |
| Palabra clave: | BACULOVIRUS RABIES VIRUS GLYCOPROTEIN POLH PSEL PROMOTER SF9 CELLS https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
| Sumario: | 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 signifcantly higher than that achieved with the standard polh promoter alone. The protein was recovered from the cell lysates and easily purifed in only one step by metal ion afnity chromatography, with a yield of 95% and a purity of 87%. Finally, GE was successfully used in an assay to detect specifc antibodies in serum samplesderived from rabies-vaccinated animals. The efcient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein. |
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