Recombinant EV71 Anti-EV71 Capsid protein VP1 Antibody (D5) (CAT#: PABC-219)

Figure 1 Binding specificities of the MAbs.

Three forms of EV71 particles (EV71-F, EV71-E, and EV71 VLP) andtwo inactivated viruses (CA16 and CA10) were examined for their reactivity tothe MAbs by ELISA.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.

Figure 2 Abilities of D5, H7, and C4 to neutralize the EV71 G082 strain.

The neutralization IC50 for each MAb is shown.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.

Figure 3 Inhibition of D5-mediated neutralization by the selected peptides.

VP1-derived peptides 1, 5, as well as an irrelevant control peptide Fd, were used for incubation with D5 or for 1h at 37°C. The peptide-MAb mixtures were subjected to neutralization testing. Neutralization inhibition by the peptides was determined using an MTT method.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.

Figure 4 D5 blocks virus attachment to susceptible cells.

Different amounts of D5 was mixed with 9×106 TCID50s of EV71 G082, and then the mixture was incubated at 37°C for 1 h before it was added to RD cells. The cells were incubated at 4°C for 1h to allow virus attachment and then washed 3 times with cold PBS to remove unbound virus. The cell-attached virus was analyzed by Western blotting.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.

Figure 5 Localization of EV71 and MAbs revealed by confocal microscopy.

EV71 F particles were preattached to Vero cells, and then the cells were treated with D5. The cells were then incubated at 37°C for 120 min.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.

Figure 6 D5 treatment interferes with EV71 binding to SCARB2 and PSGL-1.

Different amounts of D5 was mixed with EV71 G082 and then incubated at 37°C for 1 h. Subsequently, the antibody-virus mixtures were incubated with immobile SCARB2 on agarose beads. After washing to remove unbound virus, the agarose beads were pre-cipitated by centrifugation, and the binding of virus was detected by Westernblotting with a virus-specific antibody.

Ku, Z., Ye, X., Shi, J., Wang, X., Liu, Q., & Huang, Z. (2015). Single neutralizing monoclonal antibodies targeting the VP1 GH loop of enterovirus 71 inhibit both virus attachment and internalization during viral entry. Journal of virology, 89(23), 12084-12095.