Recombinant Human Anti-EBV BLLF1 Antibody (72A1) (CAT#: PABZ-138)

Figure 1 Immunofluorescence

Lytically induced 293-BLLF1-KO cells do not show any expression of the BLLF1 gene product. 293-B95.8/F cells carrying wild-type EBV DNA as a positive control and 293-BLLF1-KO cells carrying BLLF1-negative DNA were induced with BZLF1, harvested 3 days after induction, and fixed in pure acetone. Expression of gp125 and BLLF1 was detected by immunostaining using a monoclonal antibody directed either against gp125 or against the BLLF1 protein. In a second step, staining was visualized by incubating the cells with a goat anti-mouse IgG antibody conjugated with the Cy-3 fluorochrome. 293-B95.8/F cells show positive staining for gp125 (A) and BLLF1 (B), whereas induced 293-BLLF1-KO cells only show expression of gp125 (C) and are negative for the BLLF1 protein (D).

Janz, A., Oezel, M., Kurzeder, C., Mautner, J., Pich, D., Kost, M., ... & Delecluse, H. J. (2000). Infectious Epstein-Barr virus lacking major glycoprotein BLLF1 (gp350/220) demonstrates the existence of additional viral ligands. Journal of virology, 74(21), 10142-10152.

Figure 2 Blocking experiments inhibit EBV activation of NF-B in HEK293 cells transfected with TLR2.

(A) TLR2/NF-κB-transfected HEK293 cells were preincubated for 1 h at 37°C with blocking anti-TLR2 antibodies (6 μg/ml) prior to mock stimulation or stimulation with infectious EBV for 8 h, and the luciferase assay was performed. (B) Transiently transfected HEK293 cells were mock stimulated or stimulated with infectious EBV or with EBV particles preincubated with F(ab')₂ fragments of 72A1 monoclonal antibodies (37°C, 60 min) prior to cell treatment. In both panels A and B, assays were performed in triplicate, and results are expressed in relative light units (RLU) per microgram of protein (mean plus standard error of the mean [error bar]). Values that were significantly different (P < 0.05) from the value for the mock control (M) are indicated (*). NS, not stimulated; BSA, bovine serum albumin.

Gaudreault, E., Fiola, S., Olivier, M., & Gosselin, J. (2007). Epstein-Barr virus induces MCP-1 secretion by human monocytes via TLR2. Journal of virology, 81(15), 8016-8024.

Figure 3 Antibodies generated in mice immunized with VLPs from 293-VII+ cells neutralize infectious EBV.

GFP-positive EBV 2089 stocks were preincubated for 30 min with sera from mice immunized with VLPs or exosomes and subsequently used to infect human primary B cells at a calculated multiplicity of infection of 0.1. Forty-eight hours later, the number of GFP-expressing infected cells was determined by flow cytometry. The neutralizing anti-gp350 antibody 72A1 was used as a positive control at two different concentrations.

Ruiss, R., Jochum, S., Wanner, G., Reisbach, G., Hammerschmidt, W., & Zeidler, R. (2011). A Virus-Like Particle-Based Epstein-Barr Virus Vaccine. Journal of Virology, 85(24), 13105–13113.

Figure 1 Recombinant Human Anti-EBV BLLF1 Antibody (PABZ-138) in SDS-PAGE

SDS-PAGE analysis of PABZ-138 in non-reduced (Lane 1) and reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

Figure 2 Recombinant Human Anti-EBV BLLF1 Antibody (PABZ-138) in SEC-HPLC

The purity of PABZ-138 was greater than 95% as determined by SEC-HPLC.

Figure 3 Recombinant Human Anti-EBV BLLF1 Antibody (PABZ-138) in ELISA

ELISA analysis of PABZ-138 was performed by coating with Epstein-Barr virus (Herpesvirus 4) EBV Glycoprotein gp350 / EBV GP350 Protein (His Tag).

The secondary antibody: HRP-anti-Human IgG

Figure 4 Recombinant Human Anti-EBV BLLF1 Antibody (PABZ-138) in in WB

Western blot analysis of PABZ-138 was performed with Epstein-Barr virus (Herpesvirus 4) EBV Glycoprotein gp350 / EBV GP350 Protein (His Tag).
PABZ-138 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-anti-Human IgG (H+L)

Figure 5 Recombinant Human Anti-EBV BLLF1 Antibody (PABZ-138) in DB

Dot Blot analysis of PABZ-138 was performed by coating with Epstein-Barr virus (Herpesvirus 4) EBV Glycoprotein gp350 / EBV GP350 Protein (His Tag).

The secondary antibody: HRP-anti-Human IgG