Monkey Anti-HIV-1 Env Recombinant Antibody (clone JR4) (CAT#: PABL-148)

Figure 1 The four panels illustrate the enhanced exposure of ADCC Ab epitopes in gpl20-M2/VRC01 ScFv fusion proteins.

Figure 2 FRET histograms of donor (A488)-labeled Fabs, acceptor (A568)-labeled Fabs, and full-length single-chain gp120BaL-CD4 complex (FLSC) in solution as determined by the FRET-FCS approach.

Data in panels A and B indicate ∼20% FRET efficiency for A488-labeled C11 Fab and A568-labeled A32 Fab or N60-i3 Fab bound to FLSC, respectively. Panels D and E do not show detectable FRET signals for A488-labeled A32 Fab and A568-labeled N60-i3 Fab or JR4 Fab in the presence of FLSC in solution. From the FCS measurements, the diffusion coefficient of dye-labeled Fab is ∼83 μm2/s. This diffusion has been significantly decreased upon binding two Fabs to FLSC. The diffusion coefficient for this immune complex is ∼36 μm2/s.

Gohain, N., Tolbert, W. D., Acharya, P., Yu, L., Liu, T., Zhao, P.,... & Martin, L. (2015). Co-crystal structures of antibody N60-i3 and antibody JR4 in complex with gp120 define more Cluster A epitopes involved in effective antibody-dependent effector function against HIV-1. Journal of virology, JVI-01232.

Figure 3 MAb A32/C11 competition of N60-i3/JR4 Fab binding as measured by a surface plasmon resonance competition assay.

MAb A32 (A and B) and MAb C11 (B, C, and D) were immobilized on a protein A chip, and FLSC antigen was loaded to form a competitor antibody-antigen complex. Various concentrations of Fab MAb were tested. N60-i3 (A and C), JR4 (B and D), and A32 (E) were then passed over the chip. (F) Sensorgrams of the concentration series are shown. Binding affinities for MAbs A32, N60-i3, and JR4 to FLSC in the absence (blue bars) and presence (red bars) of C11 IgG were calculated with the BIAevaluation software.

Gohain, N., Tolbert, W. D., Acharya, P., Yu, L., Liu, T., Zhao, P.,... & Martin, L. (2015). Co-crystal structures of antibody N60-i3 and antibody JR4 in complex with gp120 define more Cluster A epitopes involved in effective antibody-dependent effector function against HIV-1. Journal of virology, JVI-01232.

Figure 4 Representative ADCC curves for MAbs N60-i3 and JR4. ADCC assays were performed using CEM-NKr-CCR5 target cells sensitized with gp120 of HIV-1BaL isolate (A) or spinoculated with AT-2-inactivated BaL viruses (B). MAb palivizumab was included as a control.

Gohain, N., Tolbert, W. D., Acharya, P., Yu, L., Liu, T., Zhao, P.,... & Martin, L. (2015). Co-crystal structures of antibody N60-i3 and antibody JR4 in complex with gp120 define more Cluster A epitopes involved in effective antibody-dependent effector function against HIV-1. Journal of virology, JVI-01232.