Fig. 2: BinJ/JEVNSW22-prME vaccine antigen purification.
a BinJ/JEVNSW/22-prME was purified via a potassium tartrate gradient, sedimenting as a wide opalescent blue band (mature virions) or white band (immature virions). b SDS-PAGE (4–12%) analysis of gradient purified BinJ/JEVNSW/22-prME. Flavivirus structural proteins (pre-membrane (prM), capsid (C), membrane (M) and envelope (E)) are indicated. c Negative-stain TEM image of mature (upper panel, x20, 000 magnification) and immature (lower panel, x25, 000 magnification) BinJ/JEVNSW/22-prME virions. Scale bar represents 200 nm. For lower panel, example of mature virion (red arrow) and immature virion (black arrow) indicated. d ELISA curves for indicated purified anti-JEV mAbs using C6/36 fixed cells that had been infected with BinJV-based JEV chimeras GI-V, wild-type WNVKUN and MVEV or mock-infected as the antigen. e Competitive ELISA analysis using saturating concentrations of mAbs listed on the X axis and detection of binding of mAb JV-4H12. f Kd values for binding of mAbs to BinJ/JEVNSW/22-prME and the corresponding JEVNSW/22; each dot represents one mAb (mAbs are described in Table S4). Yellow, E-specific mAbs; orange, E domain II–specific mAbs; green, E domain III–specific mAbs; purple, prM and E-specific mAbs, grey, unspecified prM/E-reactive mAbs. Statistics were performed using Pearson correlations.
