Human Anti-HIV Env Recombinant Antibody (clone 10-1074) (CAT#: PABL-508)

Figure 1 Antibody binding at the surface of cells infected with two HIV-1 reference strains.

CEM-NKR cells infected with HIV-1 (NLAD8) were incubated with 15 μg/ml of anti-Env monoclonal antibodies. The levels of antibody bound on infected (Gag-high) and bystander (Gag-low and Gag-negative) cells were then evaluated by flow cytometry.

Bruel, T., Guivel-Benhassine, F., Lorin, V., Lortat-Jacob, H., Baleux, F., Bourdic, K., ... & Schwartz, O. (2017). Lack of ADCC breadth of human nonneutralizing anti-HIV-1 antibodies. Journal of virology, 91(8), e02440-16.

Figure 1 Human Anti-HIV Env Recombinant Antibody (PABL-508) in SDS-PAGE

SDS-PAGE analysis of PABL-508 in β-mercaptoethanol-reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, the different reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of PABL-508 was greater than 95% as determined by SDS-PAGE.

Figure 2 Human Anti-HIV Env Recombinant Antibody (PABL-508) in SEC-HPLC

The purity of PABL-508 was 99.4989% as determined by SEC-HPLC.

Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl

Figure 3 Human Anti-HIV Env Recombinant Antibody (PABL-508) in WB

Western blot analysis of PABL-508 was performed by loading recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein, (His tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a NC membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-508 and HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000). Chemiluminescent detection was performed.

Lane 1: Reduced antigen (0.1 μg)
Lane 2: Reduced antigen (0.3 μg)
Lane 3: Reduced antigen (0.6 μg)

Figure 4 Human Anti-HIV Env Recombinant Antibody (PABL-508) in DB

Dot Blot analysis of PABL-508 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein (His tag).

PABL-508 incubation concentration: 2 μg/mL.
HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000)

Figure 5 Human Anti-HIV Env Recombinant Antibody (PABL-508) in ELISA

ELISA analysis of PABL-508 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein, (His tag). Then blocked with BSA and incubated with PABL-508. The HRP conjugated Anti-Human IgG as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.