Figure 4 | Scientific Reports

Figure 4

From: Nanobodies mapped to cross-reactive and divergent epitopes on A(H7N9) influenza hemagglutinin using yeast display

Figure 4

Epitope footprint of H3/H7 cross-reactive nanobodies. (a) Flow cytometry histograms (FlowJo 10.4 software) showing NB37X-01 and NB37X-05 binding to displayed wild type H7-HA0 and mutants selected (b) Nb binding activity to a panel of yeast displayed HA0 mutants. Residue numbering is relative to HA2 domain. The Nbs used for epitope mapping are highlighted in blue. Commercial antibody MIA-H7-334 is included as positive control and retains binding to all mutants. Relative binding of Nbs to each displayed mutant were categorized as follows: ≤ 15% no binding (red), between 15 and 40% intermediate binding (orange) and ≥ 40% strong binding (green). *Represents retention of binding to the HA1 head domain carrying a non-relevant mutation HA1-S135A (c) yeast cells displaying the head specific Nb NB7-14 were incubated with HA treated with low and neutral pH and binding of the stem specific Nbs NB37X-01 and NB37X-05 was detected as mean fluorescent intensity (MFI). NB7-04 was a non-competing head specific nanobody control.

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