Article Title: Recombinant Fc-fusion vaccine of RBD induced protection against SARS-CoV-2 in non-human primate and mice
Figure Lengend Snippet: SARS-Cov2 RBD-Fc vaccine Design. a) Two RBD domains are fused through Fc fragment to form the Y-shaped structure via protein structure prediction server version 3.0 (left panel). RBD-Fc protein expressed from CHO cells were identified by western blot under reduced and non-reduced condition using both anti-serum from COVID-19 recovered patients and commercial antibody (right panel). b) N-glycosylation and O-glycosylation sites identified using mass spectrum (up panel). The docking between ACE-2 and RBD-Fc predicted by ZDOCK server. An overview of the glycosylation sites illustrated based on the solved complex structure of SARS-CoV-2 RBD-Fc bound to ACE2 (PDB code: 1R42). The identified sites, colored red for N-glycosylation, purple for O-glycosylation are shown as spheres and labeled. The right panel (surface representation) was generated by rotating the structure in the Left panel (cartoon representation) around a vertical axis for about 90° (lower panel). c) The real-time binding profile between our purified RBD-Fc protein and ACE2 characterized by SPR Biacore. d e) Balb/C mice were immunized with RBD-his or RBD-Fc (10ug/mouse) in the presence of aluminum at d0 and d14. Two weeks post last vaccination the serum were collected ELISA assay shows the SARS-CoV-2 RBD-specific IgG titers (d). SARS-CoV-2 neutralization assay shows the NT50 (e).
Article Snippet: The cell culture supernatants were collected and analyzed by western blot analysis using a commercial antibody (Sino Biological Inc. Beijing, China) against SARS-CoV-2 RBD.
Techniques: Western Blot, Labeling, Generated, Binding Assay, Purification, SPR Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Neutralization