Human Anti-Ebola virus glycoprotein Recombinant Antibody (Clone 114) (CAT#: PABS-0003)

Figure 1 Immunoprecipitation (IP) of soluble GP ectodomain containing or lacking the mucin-like domain (GPΔMuc) by mAb100, mAb114 or isotype control.

Binding and input were analyzed using immunoblotting for GP1. * Represents a GP1 degradation product present only in mucin-containing GP. (n=3, representative image shown)

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.

Figure 2 GPΔMuc was incubated with the indicated antibodies followed by cleavage at pH 5.5 by Cat L at 37 °C.

Samples were removed at 5 minute intervals and analyzed by immunoblot for GP1. (n=3, representative image shown)

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.

Figure 3 Immunoprecipitation of GPΔMuc and GPTHL by the indicated antibodies.

Samples were analyzed by immunoblot for GP1. (n=3, representative image shown)

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.

Figure 4 Class averages of single particles from negative-stain electron micrographs of Fab114 in complex with GPΔMuc and GPTHL.

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.

Figure 5 Binding kinetics of GPΔMuc or GPTHL with Fab114, 13C6 or monomeric domain C of NPC1 (NPC1-dC) at the indicated pH as measured by biolayer interferometry.

Equilibrium dissociation constants (KD) are plotted on a negative log scale. * indicates no binding. (n=2, representative experiment shown)

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.

Figure 6 Inhibition of NPC1-dC binding to GPTHL by competitor proteins (NPC1-dC) or antibodies (mAb100, mAb114, 13C6, KZ52, isotype control) was determined by biolayer interferometry.

Dashed line represents 60% inhibition of binding. (n=3, representative experiment shown)

Misasi, J., Gilman, M. S., Kanekiyo, M., Gui, M., Cagigi, A., Mulangu, S., ... & McLellan, J. S. (2016). Structural and molecular basis for Ebola virus neutralization by protective human antibodies. Science, 351(6279), 1343-1346.