Recombinant Human Anti-HIV-1 gp120 Antibody (45-46M2) (CAT#: PABZ-169)

Figure 1 Comparison of neutralization potencies of 45-46m bNAb mutants.

(A) Summary of 45-46m mutants. (B) Coverage curves showing the cumulative frequency of IC₅₀ values up to the concentration shown on the x axis (plot of the percent of viral strains [y axis] from a panel of 118 strains that were neutralized at a given IC₅₀ cutoff [x axis]). A vertical line at 0.1 µg/ml designates a theoretical desired potency for a therapeutic reagent. (C) Table showing IC₅₀ values (µg/ml) for NIH45-46, NIH45-46^G54W, 45-46m2, and 45-46m7 against 28 strains that are resistant to or poorly neutralized by NIH45-46. Strains marked in blue have an altered N/DNGG motif. IC₅₀s were derived from curves generated from data points obtained in duplicate or triplicate.

Diskin, R., Klein, F., Horwitz, J. A., Halper-Stromberg, A., Sather, D. N., Marcovecchio, P. M., ... & Stamatatos, L. (2013). Restricting HIV-1 pathways for escape using rationally designed anti–HIV-1 antibodies. Journal of Experimental Medicine, 210(6), 1235-1249.

Figure 2 Neutralization of highly resistant viral clones from patient VC10042.

(A) Neutralization curves for NIH45-46^G54W, 45-46m2, and 45-46m7 against 10 viral clones from patient VC10042 that were isolated 19 yr (first three panels) or 22 yr (remaining panels) after infection. (B) Scatter plot comparing IC₅₀ values (µg/ml) for VRC01, NIH45-46^G54W, 45-46m2, and 45-46m7 against viral clones from patient VC10042. Despite their breadth, 45-46m2 and 45-46m7 did not neutralize SIV strains (SIVmac251 and SIVsmE660; not depicted). (C) IC₅₀ values (µg/ml) for NIH45-46^G54W, 45-46m2, and 45-46m7 against viral clones from patient VC10042. The reported IC₅₀ values represent the mean of two independent experiments, each with two replicates. Significance of statistical differences: * (VRC01 and 45-46m2), P < 0.0001; ** (NIH45-46^G54W and 45-46m2), P = 0.0033; *** (45-46m2 and 45-46m7), P = 0.0274. Error bars represent standard deviation from the mean.

Diskin, R., Klein, F., Horwitz, J. A., Halper-Stromberg, A., Sather, D. N., Marcovecchio, P. M., ... & Stamatatos, L. (2013). Restricting HIV-1 pathways for escape using rationally designed anti–HIV-1 antibodies. Journal of Experimental Medicine, 210(6), 1235-1249.

Figure 3 SPR comparisons of the binding of gp120 to NIH45-46, NIH4546G54W, and 45-46m2 Fabs.

(A) Sensograms (orange curves) were recorded for the interactions of injected 93TH057 gp120 produced in insect (Hi5) and mammalian (HEK293) cells over immobilized Fabs derived from the indicated antibodies in a twofold dilution series ranging from 500 to 31 nM. Kinetic constants (kₐ, k_d) were derived from globally fitting the association and dissociation phases using a 1:1 binding model (black curves) and affinities were calculated as K_D = k_d/kₐ. Residual plots (blue) within each sensogram describe the fit of the model to the data. Each binding experiment was conducted twice; once using gp120 produced in insect cells and once using gp120 produced in mammalian cells. (B) SPR measurements of 500 nM injected 93TH057 gp120 over the indicated immobilized Fabs. Each curve was normalized to its R_max. The gray and white shaded areas designate the association and dissociation phases, respectively.

Diskin, R., Klein, F., Horwitz, J. A., Halper-Stromberg, A., Sather, D. N., Marcovecchio, P. M., ... & Stamatatos, L. (2013). Restricting HIV-1 pathways for escape using rationally designed anti–HIV-1 antibodies. Journal of Experimental Medicine, 210(6), 1235-1249.

Figure 4 Levels of neutralizing mAbs in the plasma of passively transferred macaques.

The concentrations of the indicated mAbs in plasma at various times after administration were determined by ELISA using recombinant HIV-1 gp120. Plasma antibody levels were measured in the animals described in Table 2. Means and SDs are shown.

Shingai, M., Donau, O. K., Plishka, R. J., Buckler-White, A., Mascola, J. R., Nabel, G. J., ... & Diskin, R. (2014). Passive transfer of modest titers of potent and broadly neutralizing anti-HIV monoclonal antibodies block SHIV infection in macaques. Journal of Experimental Medicine, 211(10), 2061-2074.