Fig. 2: Design and in vitro characterization of RSV-F UFCR1 series.
SEC profile (left top), DSC profile (left bottom), representative 2D classification images (middle, or right when no 3D models are shown), and 3D reconstruction from nsEM analysis (right) for (a) UFCR1, (b) UFCR1-P2, (c) UFCR1-P2-NQ, and (d) UFCR1-P2-L2. All UFCR1 constructs were transiently expressed in 25 ml of ExpiCHO cells and purified using a D25 antibody column. The trimer (T) peak is marked on the profile. e The nsEM analysis of UFCR1-P2-NQ in the presence of prefusion-specific antibody D25 (left panel) or postfusion-specific antibody ADI-14359 (right panel). Each panel shows the ribbon model of the RSV-F/antibody complex (left), representative 2D classification images (middle), and 3D reconstruction (right). The 2D classes corresponding to prefusion-closed trimers (either ligand-free or antibody-bound) are circled in blue, and a prefusion RSV-F trimer (PDB ID: 4JHW) was used for structural fitting into the nsEM densities. f ELISA-derived EC50 (µg/ml) values of four UFCR1 constructs binding to 10 antibodies, as in Fig. 1e. g BLI-derived antigenic profiles of four UFCR1 constructs binding to 10 antibodies. Sensorgrams were obtained using the same protocol as in Fig. 1f and are shown in Fig. S2e. The matrix of peak values at the highest antigen concentration is shown, as in Fig. 1f.
