Human Anti-HIV-1 gp41 Recombinant Antibody (clone 3BC315) (CAT#: PABL-188)

Figure 1 Neutralization of wild type and L619Y mutant JR-FL (left) and BG505 (right) pseudoviruses by 3BC176 and 3BC315 IgG, measured in a TZM-bl neutralization assay.

Lee, J. H., Leaman, D. P., Kim, A. S., De La Peña, A. T., Sliepen, K., Yasmeen, A.,... & Klein, F. (2015). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. Nature communications, 6, 8167.

Figure 2 Binding of 3BC176 and 3BC315 to wild type and W623A BG505 SOSIP.664293T measured by ELISA. This mutation in the heart of the epitope clearly reduces 3BC176 and 3BC315 binding to the SOSIP trimer. Binding of the trimer-specific antibody PGT145 confirms that the W623A mutant does not adversely affect the quaternary structure of the trimer.

Lee, J. H., Leaman, D. P., Kim, A. S., De La Peña, A. T., Sliepen, K., Yasmeen, A.,... & Klein, F. (2015). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. Nature communications, 6, 8167.

Figure 3 SPR analysis of sequential antibody binding to BG505 SOSIP.664293T trimer.

The sequential-injection curve is colour-coded with the label of its first Ab. The single-injection curve for the second Ab in the sequential injection is superimposed on its corresponding curve in the second injection in a different colour. The curves are displayed in the same order as shown in the matrix to the right. The matrix diagram gives the binding of the second Ab in a sequential injection relative to its binding when injected alone (% of plateau colour-coded as shown in key at the bottom).

Lee, J. H., Leaman, D. P., Kim, A. S., De La Peña, A. T., Sliepen, K., Yasmeen, A.,... & Klein, F. (2015). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. Nature communications, 6, 8167.

Figure 4 JR-FL and BG505 virions were incubated at 37 °C alone or with 20 μg ml−1 sCD4, 3BC176, 3BC315, PGT151 or PG9 Fabs for noted time points.

Virions were then pelleted and gp120 shed into the cell culture supernatant (upper panels) or associated with virus (lower panels) was detected using ELISA. The % of increase in gp120 (upper panels) and the % of gp120 remaining with the virus (lower panels) are relative to the untreated control. The scales are not the same in all figures. In both measurements, gp120 shedding is observed when the virus is treated with sCD4 (control), 3BC176 or 3BC315, but not when treated with PG9 or PGT151. Error bars indicate s.d.

Lee, J. H., Leaman, D. P., Kim, A. S., De La Peña, A. T., Sliepen, K., Yasmeen, A.,... & Klein, F. (2015). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. Nature communications, 6, 8167.

Figure 5 Pseudovirus neutralization measured in TZM-bl assay. Graphs show percent neutralization (y axis) by increasing concentrations of 3BC176 or 3BC315 (x axis) of wt SF162 and mutants lacking the V1 or the V2 loop (SF162 DV1 and SF162 DV2, respectively). (D) IC50 of 3BC176 and 3BC315 neutralization of SF162 wt and 11 SF162 pseudoviruses carrying single mutations at different glycosylation sites or the K160N mutatio.n Position of mutated glycosylation site (x axis) according to HXBc2. For both antibodies, the fold increase or decrease of the IC50 values are visualized.

Klein, F., Gaebler, C., Mouquet, H., Sather, D. N., Lehmann, C., Scheid, J. F.,... & Poignard, P. (2012). Broad neutralization by a combination of antibodies recognizing the CD4 binding site and a new conformational epitope on the HIV-1 envelope protein. Journal of Experimental Medicine, 209(8), 1469-1479.

Figure 6 Inhibition of 3BC176 and 3BC315 binding to gp160 ΔcBaL-transfected 293T cells in the presence of sCD4 (10 µg/ml). Graphs show inhibition (in percentage) of Alexa Fluor 647–labeled 3BC176 or 3BC315 (y axis) in the presence of increasing concentrations of the indicated antibodies (x axis).

Klein, F., Gaebler, C., Mouquet, H., Sather, D. N., Lehmann, C., Scheid, J. F.,... & Poignard, P. (2012). Broad neutralization by a combination of antibodies recognizing the CD4 binding site and a new conformational epitope on the HIV-1 envelope protein. Journal of Experimental Medicine, 209(8), 1469-1479.