Mouse Anti-EBV gH/gL Recombinant Antibody (clone CL59) (CAT#: PABC-523)

Figure 1 CL59 inhibition of membrane fusion.

Inhibition of fusion activity with purified CL59 mAb (lighter color shade) and Fab (darker color shade) using epithelial cells. The x-axis indicates the amount of purified antibody as final concentration in nanomoles.

Sathiyamoorthy, K., Jiang, J., Möhl, B. S., Chen, J., Zhou, Z. H., Longnecker, R., & Jardetzky, T. S. (2017). Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies. Proceedings of the National Academy of Sciences, 114(41), E8703-E8710.

Figure 2 gH-null virus can infect 293 cells expressing gHgL.

gH-null virus can infect 293 cells expressing gHgL. 293 cells were transfected with plasmids expressing gHgL, and 36 h later they were infected with Akata-gH-null virus. At 48 h postinfection, glycoprotein expression was detected using monoclonal antibody CL59 to gHgL and goat anti-mouse antibody conjugated to Alexa Fluor 647, and infection was monitored in terms of GFP expression.

Chesnokova, L. S., Ahuja, M. K., & Hutt-Fletcher, L. M. (2014). Epstein-Barr virus glycoprotein gB and gHgL can mediate fusion and entry in trans, and heat can act as a partial surrogate for gHgL and trigger a conformational change in gB. Journal of virology, 88(21), 12193-12201.

Figure 3 gHgL mutant surface expression and conformation was monitored using anti-gHgL E1D1, CL40 and CL59 monoclonal antibodies and HL800 polyclonal serum using a cell ELISA assay.

The results are expressed as % change in antibody binding relative to wt, with most antibodies showing no changes in binding to the mutants relative to the wt gHgL (values near zero represent wt binding). E1D1 binding is selectively lost for mutations in gl residues gl:L74, glL:176 and glL:Y131. n.d., not detected. A western blot of the gi N69 glycosylation mutants detected using a polyclonal anti-gHgL antibody (rabbit) is shown adjacent to the antibody binding data, with monoclonal anti-GAPDH blots shown as a loading control. The gel shows the expected shift in molecular weight of the gl mutants due to loss of glycan at the mutated NXS motif.

Sathiyamoorthy, K., Hu, Y. X., Möhl, B. S., Chen, J., Longnecker, R., & Jardetzky, T. S. (2016). Structural basis for Epstein–Barr virus host cell tropism mediated by gp42 and gHgL entry glycoproteins. Nature communications, 7, 13557.

Figure 4 Isolated mAb Neutralizes EBV Infection in B Cells.

Serial dilutions of the indicated antibodies were evaluated for their ability to neutralize B95.8/F EBV infection of B (Raji) cells. AntigH/gL antibodies are shown in shades of blue, anti-gB antibodies are shown in shades of red, and the anti-gp350 antibody (72A1) is shown in gray. The human or murine origins of the antibodies are indicated in the legend. Data points are mean ± SD of duplicate wells. Representative curves from 2 to 6 replicates are shown.

Snijder, J., Ortego, M. S., Weidle, C., Stuart, A. B., Gray, M. D., McElrath, M. J., ... & McGuire, A. T. (2018). An antibody targeting the fusion machinery neutralizes dual-tropic infection and defines a site of vulnerability on Epstein-Barr virus. Immunity, 48(4), 799-811.

Figure 1 Recombinant Mouse Anti-EBV gH/gL Antibody (PABC-523) in SDS-PAGE

Lane M: Marker
Lane 1: PABC-523 in non-reduced condition.
Lane 2: PABC-523 in reduced condition.

Figure 2 Recombinant Mouse Anti-EBV gH/gL Antibody (CL59) (PABC-523) in SDS-PAGE

SDS-PAGE analysis of PABC-523 in non-reduced (Lane 2) and reduced (Lane 1) 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 3 Recombinant Mouse Anti-EBV gH/gL Antibody (CL59) (PABC-523) in SEC-HPLC

SEC-HPLC analysis of PABC-523 shows that the purity is more than 95%

Figure 4 Recombinant Mouse Anti-EBV gH/gL Antibody (CL59) (PABC-523) in WB

Western blot analysis of PABC-523 was performed with Epstein-Barr virus Glycoprotein H & Glycoprotein L Heterodimer Protein, Twin-Strep&His Tag.
Lane 1: Reducing antigen (0.1 μg)
Lane 2: Reducing antigen (0.3 μg)
Lane 3: Reducing antigen (0.6 μg)
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)

Figure 5 Recombinant Mouse Anti-EBV gH/gL Antibody (CL59) (PABC-523) in ELISA

ELISA analysis of PABC-523 was performed by coating with Epstein-Barr virus Glycoprotein H & Glycoprotein L Heterodimer Protein, Twin-Strep&His Tag.
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)

Figure 6 Recombinant Mouse Anti-EBV gH/gL Antibody (CL59) (PABC-523) in Dot Blot

Dot Blot analysis of PABC-523 was performed with Epstein-Barr virus Glycoprotein H & Glycoprotein L Heterodimer Protein, Twin-Strep&His Tag.
The secondary antibody: HRP Goat Anti-Mouse IgG (H+L)