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Purification and characterization of rabies virus glycoprotein expressed by S2 (Drosophila melanogaster) and recombinant Semliki Forest Virus systems

Grant number: 12/24647-0
Support Opportunities:Regular Research Grants
Duration: June 01, 2013 - May 31, 2015
Field of knowledge:Biological Sciences - Immunology
Principal Investigator:Renato Mancini Astray
Grantee:Renato Mancini Astray
Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Associated researchers:Carlos Augusto Pereira ; Elisabeth de Fatima Pires Augusto ; Soraia Attie Calil Jorge


The recombinant rabies virus glycoprotein (RVGP) has been expressed and studied in the Viral Immunology laboratory (Butantan Institute), in order to obtain an effective immunogen and potential candidate for a third-generation vaccine against rabies. Two systems have been studied: Schneider 2 cells - S2 - Drosophila melanogaster and Semliki Forest virus (SFV). The RVGP produced by these systems has been used in immunization experiments showing important immunogenic properties. In this project, we search firstly to obtain a S2 cell line expressing high amounts of RVGP with histidine tag. Then, the RVGP produced by S2 or mammalian cells infected with SFV will be purified with the aid of the histidine tag present in both constructs. The purified RVGP will be characterized physico-chemically and antigenically. Different methods of membrane preparation from RVGP expressing cells, solubilization of the recombinant RVGP from the membrane followed by purification techniques, especially ultracentrifugation, chromatography and immunoprecipitation, will be assayed. In the development of these activities, a study of mammalian cell lines most appropriate for infection with SFV will be performed analyzing the penetration ability of SFV and the RVGP production. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SANTOS, NAYARA G. L.; ROCCA, MAYRA P.; PEREIRA, CARLOS A.; VENTINI, DANIELLA C.; PUGLIA, ANA LIA P.; JORGE, SORAIA A. C.; LEMOS, MARCOS A. N.; ASTRAY, RENATO M.. Impact of recombinant Drosophila S2 cell population enrichment on expression of rabies virus glycoprotein. Cytotechnology, v. 68, n. 6, p. 2605-2611, . (12/24647-0)
PILATTI, LIVIA; MANCINI ASTRAY, RENATO; ROCCA, MAYRA PEREIRA; BARBOSA, FLAVIA FERREIRA; JORGE, SORAIA ATTIE CALIL; BUTLER, MICHAEL; DE FATIMA PIRES AUGUSTO, ELISABETH. Purification of rabies virus glycoprotein produced inDrosophila melanogasterS2cells: An efficient immunoaffinity method. BIOTECHNOLOGY PROGRESS, v. 36, n. 6, . (18/10538-1, 12/24647-0)

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