Journal: International Journal of Molecular Sciences
Article Title: Neutralizing Human Antibodies against Severe Acute Respiratory Syndrome Coronavirus 2 Isolated from a Human Synthetic Fab Phage Display Library
Figure Lengend Snippet: Characterization of anti-SARS-2 RBD immunoglobulin Gs (IgGs). ( a ) Binding analysis of five human anti-SARS-2 RBD IgGs—C12 (IgG), H1 (IgG), C2 (IgG), D12 (IgG), and F7 (IgG)—to the SARS-2 RBD and its variants (top) and the SARS-CoV-2 S1 (D614G) and other coronavirus S1 proteins (bottom), respectively. ( b ) Soluble ELISA of five serially diluted human anti-SARS-2 RBD IgGs on immobilized SARS-2 RBD surfaces to measure their apparent affinities ( EC 50 , nM). ( c ) ELISA detection for five human anti-SARS-2 RBD IgGs blocking the binding of the ACE2 protein with the SARS-CoV-2 RBD (top) and analysis of the flow cytometry for the blocking effect between the SARS-CoV-2 RBD and an ACE2-overexpressed cell (bottom). ( d ) Size-exclusion chromatography analysis of five human anti-SARS-2 RBD IgGs. The positions of the molecular mass markers, shown as kDa, on the retention time x -axis are indicated above the peaks. The data are presented as the mean ± standard error (SEM). MFI: mean fluorescence intensity; NC: negative control; *, **, and ***: p
Article Snippet: Production of SARS-CoV-2 Spike pseudovirusPlasmids encoding the SARS-CoV-2 spike protein (D614) were purchased from Sino Biological (pCMV3-SARS-CoV-2 Spike, Cat. VG40589-UT, Beijing, China).
Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Flow Cytometry, Size-exclusion Chromatography, Fluorescence, Negative Control