Fig. 1: Viral variants capable of escaping neutralization.

a, Representative neutralization curves of SNV-42, germline reverted (GR) SNV-42 and negative control DENV 2D22 to VSV/SNV determined through real-time cellular analysis using the Vero CCL-81 cell line. IC₅₀ values were calculated on the basis of a nonlinear regression and error bars denote mean ± s.d. The assay was performed three independent times with similar results. b, Affinity measurements of SNV-42 and SNV-42^GR for binding to SNV Gn^H ectodomain, measured by bio-layer interferometry. Representative curves and K_D values are shown for SNV-42^GR, while the K_D value for SNV-42 could not be determined because the K_off could not be measured. Dashed line indicates dissociation step at 300 s. c, Representative neutralization curves of SNV-42, SNV-42^GR, positive control (oligoclonal mix of SNV-reactive antibodies) and DENV 2D22 to mutant VSV/SNV viruses. Error bars denote mean ± s.d. The assay was performed three independent times with similar results. d, SNV-42 binding in the presence of SNV M-segment mutant constructs determined by flow cytometry. The percent binding (% WT) of each mAb to the mutant constructs was compared to the WT SNV construct. An oligoclonal mix of SNV-reactive antibodies was included to control for expression of each mutant construct. The data are shown as mean ± s.d.; from left to right, n = 9, 9, 9, 9, 9, 9, 6, 9, 9, 9, 9, 9, 9 and 6 technical replicates. The assay was performed three to four independent times with similar results. e, Top view of escape mutants mapped to the ANDV Gn/Gc spike (PDB: 6ZJM). The blue residues designate escape mutants. Gn is shown in white and Gc is shown in grey. All numbering for SNV sequences was based on GenBank KF537002.1.