Fig. 1: Isolation and characterization of antibodies that bind to NiV F.

A Schematic of immunization of CB6F1/J mice. Black arrows with syringes denote times of immunization, the red arrows denote the times of serum harvest or spleen harvest. B Pie chart representing mouse paired heavy-light chain sequences recovered from B cells of mice immunized with prefusion NiV F. FACS gating/sorting strategies are shown in Supplementary Fig. 1F, G and described in the Methods Section. C Representative 2D class averages from negative stain EM of prefusion NiV F, alone and complexed with Fabs. Numbers of particles in each class are listed in parentheses below each image. Details regarding the numbers of micrographs are included in the Methods Section. Numbers of particles in each class are shown in parentheses below each image. Asterisks indicate antibodies that were shown to be neutralizing in the pseudovirus neutralization assay (Fig. 1D). D Neutralization of pseudotyped virus. Error bars show the standard deviation of the mean for three technical replicates. Data is representative of two independent experiments. 2C2 is a non-neutralizing antibody. E Competition binding experiment, as measured by biolayer interferometry. Competitor antibodies are listed on the left by row, analyte antibodies are listed on the top by column. The percent competition is indicated by intensity of shading, from white (<60%) to red (100%). A value less than 60% is considered to not compete. Thicker boxes along the diagonal indicate self-antibody competition controls. Larger competition binding matrix is shown in Supplementary Figure 1H, highlighted here is the neutralizing antibodies competing with one another. Competition binding among neutralizing antibodies is representative of two independent experiments. F Diagram of the structure of prefusion NiV F. One protomer is shown as a ribbon diagram, and the remaining two protomors are shown as simulated gray traces at ~5 Å resolution. A primary sequence diagram is shown at the right. Domain 1 (D1/lateral) is colored orange, domain 2 (D2/basal) is colored red and domain 3 (D3/apical) is colored yellow. The heptad repeat is colored brown.