Recombinant Mouse Anti-VACV A14 Antibody (HE6) (CAT#: FAMAB-0337CQ)

Figure 1 Immunogold staining of purified VACV MV with anti-VACV antibodies

Sucrose-gradient purified virions were incubated with the indicated antibodies at 2 μg/ml followed by incubation with secondary gold-conjugated antibodies. The virions were stained with uranyl-acetate and visualized with an electron microscope.

Meng, X., Kaever, T., Yan, B., Traktman, P., Zajonc, D. M., Peters, B., ... & Xiang, Y. (2018). Characterization of murine antibody responses to vaccinia virus envelope protein A14 reveals an immunodominant antigen lacking of effective neutralization targets. Virology, 518, 284-292.

Figure 2 Neutralization of VACV MV by A14 mAbs

Sucrose-gradient purified VACV mature virions were incubated with the indicated concentration of purified antibodies in the presence or absence of rabbit complement for 1 hr at 4°C. The mixture was then added to monolayers of BS-C-1 cells, and the inoculum was removed after one hour. The number of plaques that appeared after 2 days was enumerated. The number of plaques obtained under the indicated condition as the percentage to the number of plaques from untreated inoculums is shown. The average and standard deviation are from three independent inoculums.

Meng, X., Kaever, T., Yan, B., Traktman, P., Zajonc, D. M., Peters, B., ... & Xiang, Y. (2018). Characterization of murine antibody responses to vaccinia virus envelope protein A14 reveals an immunodominant antigen lacking of effective neutralization targets. Virology, 518, 284-292.

Figure 3 Protection of SCID mice from intravenous VACV challenge by anti-A14 mAbs

Groups of six SCID mice were treated with either 400 μL PBS or 400 μL of the indicated antibodies at a concentration of 0.25 mg/mL (=100 μg) on day −1. PBS/antibodies were injected at 200 μL intraperitoneal and 200 μL retro-orbitally. Then, mice were challenged with 1 x 105 PFU (200 μL) of VACV ACAM2000 by the retro-orbital route on day 0. (A) Body weights. (B) Body weight loss at day 58. (C) Survival. (D) Clinical scores over time.

Meng, X., Kaever, T., Yan, B., Traktman, P., Zajonc, D. M., Peters, B., ... & Xiang, Y. (2018). Characterization of murine antibody responses to vaccinia virus envelope protein A14 reveals an immunodominant antigen lacking of effective neutralization targets. Virology, 518, 284-292.

Figure 4 Mapping the epitopes of A14 mAbs.

Further define the 8C6 epitope by ELISA of A14 mutants. 293T cells were transfected with plasmids encoding A14 alleles under the control of a VACV promoter and subsequently infected with an IPTG-inducible A14 mutant VACV (VACV-iA14) either in the absence of IPTG. The cell lysates were used to coat a microtiter plate, and ELISA was performed with either 8C6 or HE6. The A14 plasmids are named after the A14 residues expressed (12-75, 12-90) or A14 residues substituted with alanines (26–30->A, 32–35->A, 39–44->A).

Meng, X., Kaever, T., Yan, B., Traktman, P., Zajonc, D. M., Peters, B., ... & Xiang, Y. (2018). Characterization of murine antibody responses to vaccinia virus envelope protein A14 reveals an immunodominant antigen lacking of effective neutralization targets. Virology, 518, 284-292.