Recombinant Alpaca Anti-VSV N Single Domain Antibody (PABC-564) (CAT#: PABC-564)

Figure 1 VHH binding to N and the N/P complex.

Biotinylated, N-specific VHHs, and a control VHH (aNP-VHH1, influenza A nucleoprotein specific) were immobilized on streptavidin beads and N alone, or N in complex with P was added to the beads. Bound protein was eluted and subjected to 10% SDS–PAGE (75:1 acrylamide/bis-acrylamide) and Coomassie staining. A representative gel from three independent experiments is shown.

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.

Figure 2 Inhibitory properties of VSV N‐specific VHHs

VSV N-specific VHHs differentially cope with increasing doses of VSV. A549 cells expressing VHHs in a doxycycline (Dox)‐inducible manner were seeded 24 h before VSV infection. VHH expression was induced (gray to black bars) with Dox, or cells were left untreated (white bars). Each cell line was infected with increasing amounts of VSV‐GFP (MOI = 0–320).

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.

Figure 3 Inhibitory properties of VSV N‐specific VHHs

Quantification of VHH expression levels in A549 cells. Cells were seeded, and HA‐tagged VHH expression was induced with Dox.

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.

Figure 4 VHH binding to N and the N/P complex.

Biotinylated, N-specific VHHs, and a control VHH (αNP‐VHH1, influenza A nucleoprotein specific) were immobilized on streptavidin beads and N alone, or N in complex with P was added to the beads.

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.

Figure 5 Band intensities of the immunoprecipitated N and coimmunoprecipitated P were quantified, and the relative amount of P compared to N is displayed.

Band intensities of the immunoprecipitated N and coimmunoprecipitated P were quantified, and the relative amount of P compared to N is displayed.

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.

Figure 6 VHH binding to VSV N escape variants

Indicated, HA-tagged VHHs and VSV N (WT or mutated versions) fused to Renilla luciferase, or influenza A NP fused to Renilla luciferase were transiently coexpressed in 293T cells. 96-well plates coated with anti-HA antibody to capture the VHHs were incubated with cell lysates. Activity of the copurified luciferase was measured. Emitted light was normalized to luciferase activity in the lysate.

Hanke, L., Schmidt, F. I., Knockenhauer, K. E., Morin, B., Whelan, S. P., Schwartz, T. U., & Ploegh, H. L. (2017). Vesicular stomatitis virus N protein‐specific single‐domain antibody fragments inhibit replication. EMBO reports, 18(6), 1027-1037.