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