Human Anti-HIV-1 gp41 Recombinant Antibody (clone 4E10) (CAT#: PABL-192)

Figure 1 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 2 Binding of MPER-specific IgA on SupT1/LAI-infected cells. Number of IgA+/p24+ SupT1 cells was analyzed by flow cytometry. 4E10 was used as positive control and labeling with anti-p24+ + anti-IgA-PE as negative control. The ratio of the mean fluorescence of IgA+/p24+ SupT1 cells on the mean fluorescence of IgA2/p24+ SupT1 cells is indicated. Uninfected cells were used as negative control.

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 3 Competition assay for C6.11 and F4.30 binding to gp120c41 in the presence of 2F5 or 4E10. gp120c41-coated wells were preincubated with 2F5 or 4E10 at different concentrations (0–20 mg/ml). C6.11 and F4.30 were used at 5 mg/ml. Assays were performed in triplicate.

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 4 Kinetic analysis of the interaction between MPER-specific IgA and recombinant gp120c41. C6.11 (5–80 nM), F4.30 (0.5–16 nM), and 4E10 (0.5–8 nM) were injected over 8200 RU of immobilized gp120c41 in PBS containing 0.005% surfactant P20.

The binding signals shown were obtained by double referencing (subtraction of the reference surface and of buffer blank injection signals). Fits are shown as dotted lines and were obtained by global fitting of the data using a 1:1 Langmuir binding model.

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 5 Screening of the neutralizing activity of MPER-specific IgA on COT6.15 pseudoviruses and COT6.15 mutants.

MPER sequence is shown in each panel. For each mAb and mutant, the value of SD is calculated for the neutralization from two independent experiments realized in triplicate. The critical residues for neutralization are indicated in red.

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 6 Interactions of 10E8 with lipid bilayers.

A liposome flotation assay was used to detect binding to HIV-1-like membranes at 37°C by various antibodies, i.e., 17b, 4E10, 10E8, 10E8-F100aA, and 10E8-W100bA. Seven fractions throughout the sucrose gradient (from top [t] to bottom) after centrifugation were collected and analyzed by Western blotting. The heavy and light chains of antibodies are indicated by H and L, respectively.

Chen, J., Frey, G., Peng, H., Rits-Volloch, S., Garrity, J., Seaman, M. S., & Chen, B. (2014). Mechanism of HIV-1 neutralization by antibodies targeting a membrane-proximal region of gp41. Journal of virology, 88(2), 1249-1258.

Figure 1 Human Anti-HIV-1 gp41 Recombinant Antibody (clone 4E10) in SDS-PAGE analysis.

SDS-PAGE analysis of PABL-192 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 4E10) in HPLC analysis.

Result of HPLC analysis of PABL-192 shows the purity is>85%.