Purification of rabies virus glycoprotein produced in Drosophila melanogaster S2 cells: An efficient immunoaffinity method |
| |
| Authors: | Livia Pilatti PhD Renato Mancini Astray Mayra Pereira Rocca Flavia Ferreira Barbosa Soraia Attie Calil Jorge Michael Butler Elisabeth de Fátima Pires Augusto |
| |
| Affiliation: | 1. Science and Technology Institute, Federal University of São Paulo (UNIFESP), São José dos Campos, Brazil;2. Viral Immunology Laboratory, Butantan Institute, São Paulo, Brazil;3. Medical School, São Paulo State University (UNESP), Botucatu, Brazil;4. National Institute for Biotechnology Research and Training (NIBRT), Dublin, Ireland |
| |
| Abstract: | Most rabies vaccines are based on inactivated virus, which production process demands a high level of biosafety structures. In the past decades, recombinant rabies virus glycoprotein (RVGP) produced in several expression systems has been extensively studied to be used as an alternative vaccine. The immunogenic characteristics of this protein depend on its correct conformation, which is present only after the correct post-translational modifications, typically performed by animal cells. The main challenge of using this protein as a vaccine candidate is to keep its trimeric conformation after the purification process. We describe here a new immunoaffinity chromatography method using a monoclonal antibody for RVGP Site II for purification of recombinant rabies virus glycoprotein expressed on the membrane of Drosophila melanogaster S2 cells. RVGP recovery achieved at least 93%, and characterization analysis showed that the main antigenic proprieties were preserved after purification. |
| |
| Keywords: | Drosophila melanogaster S2 cells immunoaffinity purification insect cells rabies virus glycoprotein |
|
|
