Human Anti-HIV-1 Env Recombinant Antibody (clone PGDM1400) (CAT#: PABL-151)

Figure 1 Neutralization profiles of PGDM1400 and CAP256-VRC26.25 against a multi-clade panels of 208 pseudoviruses (A) and highlighting specifically the activity of both bNAbs against clade C pseudoviruses (B)

(A) Each bar represents the IC80 (μg/ml) of PGDM1400 (upper graph) or CAP256-VRC26.25 (lower graph) against a single virus. Viruses are ranked according to increasing IC80 values. Bars reaching the dotted line represent IC80 values>50 μg/mL. The red bars highlight the IC80 values for PGDM1400 (upper graph) or CAP256-VRC26.25 (lower graph) against SHIV-325c. (B) IC80 values for PGDM1400 and CAP256-VRC26.25 against SHIV-325c (red dot) and against the clade C pseudoviruses included in panel (A). The horizontal bars represent the mean of IC80 values of neutralizable pseudoviruses.

Julg, B., Tartaglia, L. J., Keele, B. F., Wagh, K., Pegu, A., Sok, D.,... & Joyce, M. G. (2017). Broadly neutralizing antibodies targeting the HIV-1 envelope V2 apex confer protection against a clade C SHIV challenge. Science translational medicine, 9(406), eaal1321.

Figure 2 Neutralization of SHIV-325c by CAP256-VRC26.25, PGDM1400 and PG9

Neutralization was measured using replication competent challenge stock SHIV-325c infection of TZM-bl cells. All 3 antibodies showed 100% neutralization of SHIV-325c with no evidence of a plateau effect below 100% neutralization.

Julg, B., Tartaglia, L. J., Keele, B. F., Wagh, K., Pegu, A., Sok, D.,... & Joyce, M. G. (2017). Broadly neutralizing antibodies targeting the HIV-1 envelope V2 apex confer protection against a clade C SHIV challenge. Science translational medicine, 9(406), eaal1321.

Figure 3 Protective efficacy of PGDM1400 and CAP256-VRC26.25-LS against SHIV-325c in rhesus macaques

Plasma viral RNA (log RNA copies/mL) are shown for animals that received placebo control (A), 2 mg/kg PGDM1400 (B), 0.4 mg/kg PGDM1400 (C), 0.08 mg/kg PGDM1400 (D), 2 mg/kg CAP256-VRC26.25-LS (E), 0.4 mg/kg CAP256-VRC26.25-LS (F), or 0.08 mg/kg CAP256-VRC26.25-LS (G). All placebo controls became infected. Protection was observed in all animals that received CAP256-VRC26.25-LS at all doses. 1/5 animals in the PGDM1400 2 mg/kg dose group and 3/4 animals in the 0.08 mg/kg PGDM1400 dose group were infected. The assay sensitivity limit was>50 RNA copies/mL.

Julg, B., Tartaglia, L. J., Keele, B. F., Wagh, K., Pegu, A., Sok, D.,... & Joyce, M. G. (2017). Broadly neutralizing antibodies targeting the HIV-1 envelope V2 apex confer protection against a clade C SHIV challenge. Science translational medicine, 9(406), eaal1321.

Figure 4 Serum concentration of CAP256-VRC26.25-LS and PGDM1400 in antibody treated animals

bNAb concentrations were determined by ELISA. The results show average serum concentrations of 19.8 μg/ml, 3.3 μg/ml and 0.75 μg/mL on the day of challenge in the CAP256-VRC26.25-LS 2 mg/kg, 0.4 mg/kg and 0.08 mg/kg groups, respectively, and 6.9 μg/mL, 2.5 μg/mL and 0.22 μg/mL on the day of challenge in the PGDM1400 2 mg/kg, 0.4 mg/kg and 0.08 mg/kg groups, respectively.

Julg, B., Tartaglia, L. J., Keele, B. F., Wagh, K., Pegu, A., Sok, D.,... & Joyce, M. G. (2017). Broadly neutralizing antibodies targeting the HIV-1 envelope V2 apex confer protection against a clade C SHIV challenge. Science translational medicine, 9(406), eaal1321.

Figure 5 Heatmaps of IC80 values for single bNAbs and bNAb combinations respectively. Rows represent Env pseudoviruses, and columns represent single and combination bNAbs. Darker hues of red indicate more potent neutralization and grey cells indicate IC80 above threshold.

Julg, B., Tartaglia, L. J., Keele, B. F., Wagh, K., Pegu, A., Sok, D.,... & Joyce, M. G. (2017). Broadly neutralizing antibodies targeting the HIV-1 envelope V2 apex confer protection against a clade C SHIV challenge. Science translational medicine, 9(406), eaal1321.

Figure 1 Human Anti-HIV-1 Env Recombinant Antibody (PABL-151) in SDS-PAGE

SDS-PAGE analysis of PABL-151 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-151 was greater than 95% as determined by SDS-PAGE.

Figure 2 Human Anti-HIV-1 Env Recombinant Antibody (PABL-151) in SEC-HPLC

The purity of PABL-151 was 98.9773% 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-1 Env Recombinant Antibody (PABL-151) in WB

Western blot analysis of PABL-151 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-151 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-1 Env Recombinant Antibody (PABL-151) in ELISA

ELISA analysis of PABL-151 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-151. 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.