Author: Lucia Grenga; Fabrice Gallais; Olivier Pible; Jean-Charles Gaillard; Duarte Gouveia; Hélène Batina; Niza Bazaline; Sylvie Ruat; Karen Culotta; Guylaine Miotello; Stéphanie Debroas; Marie-Anne Roncato; Gérard Steinmetz; Charlotte Foissard; Anne Desplan; Béatrice Alpha-Bazin; Christine Almunia; Fabienne Gas; Laurent Bellanger; Jean Armengaud
Title: Shotgun proteomics of SARS-CoV-2 infected cells and its application to the optimisation of whole viral particle antigen production for vaccines Document date: 2020_4_17
ID: kd02dehk_10
Snippet: For the kinetic, 1x10 6 Vero cells seeded into 25 cm 2 flasks were grown to cell confluence in 5 mL DMEM supplemented with 5% FCS and 0.5% penicillin-streptomycin for one night at 37°C under 9% CO2. They were infected at two multiplicities of infection (MOI): 0.01 and 0.001. Cells were harvested at 1, 2, 3, 4, and 7 days post infection (dpi). Supernatants of SARS-CoV-2 infected cells were saved for plaque assay titration to confirm production of.....
Document: For the kinetic, 1x10 6 Vero cells seeded into 25 cm 2 flasks were grown to cell confluence in 5 mL DMEM supplemented with 5% FCS and 0.5% penicillin-streptomycin for one night at 37°C under 9% CO2. They were infected at two multiplicities of infection (MOI): 0.01 and 0.001. Cells were harvested at 1, 2, 3, 4, and 7 days post infection (dpi). Supernatants of SARS-CoV-2 infected cells were saved for plaque assay titration to confirm production of infectious viral particles. Infected Vero cells were microscopically observed for cytopathic effect (CPE) at the same time points. (Thermo Fisher Scientific) prior in-gel trypsin proteolysis performed as described in Hartmann et al. [15] .
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