Human Anti-HIV-1 gp41 Recombinant Antibody (clone 2F5) (CAT#: PABL-186)

Figure 1 Binding of gp41 cluster II MAbs to anionic phospholipids.

SPR sensograms are shown for the binding of 100 μg/ml of 126-6, 167D, 98-6, 4E10, 2F5, and 13H11 MAbs to POPC-cardiolipin (25:75) (A) or POPC-POPS (25:75) (B) liposomes captured on a Biacore L1 chip.

Dennison, S. M., Anasti, K., Scearce, R. M., Sutherland, L., Parks, R., Xia, S. M.,... & Alam, S. M. (2011). Nonneutralizing HIV-1 gp41 envelope cluster II human monoclonal antibodies show polyreactivity for binding to phospholipids and protein autoantigens. Journal of virology,85(3), 1340-1347.

Figure 2 Nonneutralizing human gp41 cluster II MAbs bind strongly to oligomeric gp140 Env proteins.

SPR sensograms are displayed for the binding of 98-6, 167D, 126-6, 2F5, and 13H11 MAbs to the ConB gp140 (A), ConS gp140 CFI (B), and JRFL gp140 (C) Env proteins immobilized on a Biacore CM5 chip.

Dennison, S. M., Anasti, K., Scearce, R. M., Sutherland, L., Parks, R., Xia, S. M.,... & Alam, S. M. (2011). Nonneutralizing HIV-1 gp41 envelope cluster II human monoclonal antibodies show polyreactivity for binding to phospholipids and protein autoantigens. Journal of virology,85(3), 1340-1347.

Figure 3 Neutralization of the 2F5-resistant HIV-1JR2 mutant pseudovirus D664A by T20 in the presence and absence of 2F5 or D50.

Virus was preincubated with different concentrations of T20 in the presence or absence of a molar excess of either 2F5 or D50 (1 μM constant throughout) and then added to U87.CD4.CCR5 cells. Luciferase activity was measured after 72 h. The sequence of T20 is shown below the graph with the 2F5 epitope indicated, as well as the approximate region to which D50 binds, according to a previous study. Control experiments showed no effect of 1 μM D50 or 2F5 on the infectivity of the JR2 mutant D664A.

Dennison, S. M., Anasti, K., Scearce, R. M., Sutherland, L., Parks, R., Xia, S. M.,... & Alam, S. M. (2011). Nonneutralizing HIV-1 gp41 envelope cluster II human monoclonal antibodies show polyreactivity for binding to phospholipids and protein autoantigens. Journal of virology,85(3), 1340-1347.

Figure 4 The Octet binding curves of wt 2F5 Mab to the three probes in solution are shown for the wt MPER peptide, the linked peptide, and the ES2 2F5-epitope scaffold. The experimental binding curves are shown in blue and the applied Languir 1:1 model fitting of the curves are shown in red. Representative binding curves of the CDRH3-length altered 2F5 variants r4 and e4 are shown below.

Guenaga, J., & Wyatt, R. T. (2012). Structure-guided alterations of the gp41-directed HIV-1 broadly neutralizing antibody 2F5 reveal new properties regarding its neutralizing function. PLoS pathogens, 8(7), e1002806.

Figure 5 Alanine scanning of the 4E10 MPER region 669–673 and effects on recognition of 4E10, wt 2F5 and the CDRH3 and W/Y variants are shown. The bars represent fold-increase or -decrease in the binding constant (KD) of the Mabs to the alanine-substituted peptides compared to KDs for the wt MPER peptide. A positive bar indicates an affinity increase and a negative bar indicates an affinity decrease. The bars are color-coded for each ligand Mab. For context, the MPER residues comprising the 2F5 and 4E10 epitopes are shown and the residues 669–673 scanned by the alanine substitutions are as indicated.

Guenaga, J., & Wyatt, R. T. (2012). Structure-guided alterations of the gp41-directed HIV-1 broadly neutralizing antibody 2F5 reveal new properties regarding its neutralizing function. PLoS pathogens, 8(7), e1002806.

Figure 6 Neutralizing activity of MPER-specific IgA1 on primary and laboratory HIV-1 strains. 4E10 and 2F5 were used as positive controls. Dose-dependent neutralization was performed within a concentration range of 0.156–10 mg/ml.

Benjelloun, F., Oruc, Z., Thielens, N., Verrier, B., Champier, G., Vincent, N.,... & Genin, C. (2016). First Membrane Proximal External Region–Specific Anti-HIV1 Broadly Neutralizing Monoclonal IgA1 Presenting Short CDRH3 and Low Somatic Mutations. The Journal of Immunology, 1600309.

Figure 1 Human Anti-HIV-1 gp41 Recombinant Antibody (clone 2F5) in SDS-PAGE analysis.

SDS-PAGE analysis of PABL-186 in reduced (Lane 1) and non-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. And non-reduced protein migrates about 180 kDa.

Figure 2 Human Anti-HIV-1 gp41 Recombinant Antibody (clone 2F5) in HPLC analysis.

HPLC analysis of PABL-186 shows the purity is>95%.