Human Anti-HIV Env Recombinant Antibody (clone PGT121) (CAT#: PABL-509)

Figure 1 ELISA analysis of PGT121 binding to monomeric gp120, which was extracted from lysed virus supernatants: (A) 92BR020, (C) 92RW020, (E) JR-FL E168K/N192A, (G) IAVI C22.

Antibodies were also tested for cell surface Env binding by flow cytometry: (B) 92BR020, (D) 92RW020, (F) JR-FL E168K/N192A, (H) IAVI C22. mAb 2G12 was included as a control (gray). ELISA values are reported as optical density at 405 nm (OD405) and flow cytometry values are reported as mean fluorescence intensity (MFI).

Sok, D., Laserson, U., Laserson, J., Liu, Y., Vigneault, F., Julien, J. P., ... & Mahan, A. (2013). The effects of somatic hypermutation on neutralization and binding in the PGT121 family of broadly neutralizing HIV antibodies. PLoS pathogens, 9(11), e1003754.

Figure 1 Human Anti-HIV Env Recombinant Antibody (PABL-509) in SDS-PAGE

SDS-PAGE analysis of PABL-509 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-509 was greater than 95% as determined by SDS-PAGE.

Figure 2 Human Anti-HIV Env Recombinant Antibody (PABL-509) in SEC-HPLC

The purity of PABL-509 was 98.7602% 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-509) in WB

Western blot analysis of PABL-509 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-509 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-509) in DB

Dot Blot analysis of PABL-509 was performed by coating with recombinant HIV-1 [HIV-1/Clade B/C (CN54)] gp120 protein (His tag).

PABL-509 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-509) in ELISA

ELISA analysis of PABL-509 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-509. 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.