Skip to main content
. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: J Immunol Methods. 2018 Jul 24;461:23–29. doi: 10.1016/j.jim.2018.07.007

Figure 3. Epitope mapping of a broadly neutralizing antibody targeting the H3 HA of influenza A virus.

Figure 3.

The mAb F045–092 was produced by co-transfection of its heavy and light chain expression vectors in 293T cells, purified and tested against the H3 HA panel. (A) The sequence changes and locations of mutations as part of the epitope are shown on the 3D structure of trimeric H3 HA (adapted from PDB ID 4WE8). Mutations that caused significant reductions (≥50%) in mAb binding are highlighted in red. Percentage of the Ab binding to the mutants compared to wild type is indicated in the brackets. The overlapped H3 antigenic sites are indicated in different colors: Site A (red), Site B (blue), and Site D (cyan). (B) Association/disassociation curves of BLI for the F045–092 binding to the wild type and mutants with increased off-rate. The biosensors loaded with recHAs were incubated with the Ab for 300 sec at the association step followed by incubation in buffer for 180 sec at the disassociation step as shown on the X-axis. The changes of thickness at the tip of biosensors caused by Ab-Ag binding are shown by the Y-axis. The binding curve for each recHA is labeled with a unique color.