Neutralization titers of <i>N</i>.<i>benthamiana</i>-derived bnAbs and control bnAbs against six Tier 2 HIV isolates (A) and against three SHIV isolates (B) using pseudovirus-based TZM-bl cells.

<p>IC50 values are color coded as indicated. Control bnAb produced in CHO (C) and VRC01 produced in HEK293 (H) cells. CHO1–31 is a pool of two CHO-1 (PG9-like) and CHO-31 (VRC01-like) bnAbs which strongly neutralizes all HIV isolates.</p

Inhibition of VRC01, 10–1074, b12 and PGT121 neutralizing activity in the TZM-bl assay against HIV RHPA4259.7 and SHIV-BaL-P4 by different macaque anti-idiotypic antibodies.

<p>Concentrations of each of the bnAb that inhibited the target virus at 50–80% was pre-incubated with or without serial dilutions of 7 different monkey plasma samples (color coded) prior to adding virus. Deviations from this “no serum” control line indicates interference from the macaque sample with neutralization of the bnAb.</p

Circulatory clearance of bnAbs in macaques at different times after i.v. injection as measured by ELISA.

<p>Data are an average of two macaques. Each pharmacokinetics study was repeated once or twice. (A) Plasma levels (μ g/ml) following injection of mVRC01 at 5mg/kg (blue) or 10mg/kg (red) and VRC01 (10mg/kg, green). The insert shows clearance of the non-mutated VRC01 in each of two macaques on a different scale. (B) Average μ g/kg of two macaques injected with b12 (5 and 7.5 mg/kg), 10E8, 10–1074, PGT121 (each at 5mg/kg). The insert shows a comparison between 10–1074 and mNIH45–46^G54W both injected at 5mg/kg (C) Comparison of clearance of PGT121 (5mg/kg) as measured by ELISA (μ g/ml) and a pseudovirus-based TZM-bl assay against the RHPA4259.7 isolate (ID50) at different days following i.v. injection into macaques.</p

Comparison of plasma levels of anti-VRC01 antibody in all macaques injected twice with VRC01 by ELISA using plant-derived VRC01 (A) and HEK293-derived VRC01 (B).

<p>The numbers of the six macaques injected (2 macaques/gp in three different studies) are indicated and the times of the bleeds are color coded.</p

Reactivity of plasma from macaques injected with different bnAbs.

<p>(A) Plasma samples from the 11 macaques injected (color coded) were collected 2–3 weeks following the second injection and idiotype specificity was demonstrated by ELISA against PGT121, 10–1074, VRC01 and NIH45–46 produced in mammalian cells. (B) Screening of 14 naïve macaques for pre-existing reactivity against PGT121, 10–1074, VRC01 and NIH45–46^G54W prior to injection.</p

Immunogenicity of bnAbs in macaques following two injections two weeks apart.

<p>Plasma samples collected before injection (PB) and at the time indicated from macaques receiving (A) VRC01 (#5191 and #5193) or b12 (#5194 and #5192) and (B) mVRC01 (#5544 and #5545), 10-1074 (#5540 and #5542) and mNIH45–46^G54W (#5541 and #5543). Each sample was assayed by ELISA against the injected bnAb and, as a control against, b12 to demonstrate anti-bnAb antibodies.</p

Western blotting analysis of purified plant-derived and CHO-derived HIV-1 env using 2G12.

<p>(lane 1∶89.6P gp140ΔCFI-KDEL (#188), lane 2∶89.6P gp140ΔCFI (#188AH) and lane 3: <b>^BAL</b>CHO gp120. 200 ng of each sample was loaded.</p

Comparison of the neutralization titers of <i>N.benthamiana</i>/p19-derived and control mAbs at different harvest times.

<p>IC50s represent the concentration at which relative luminescence units (RLUs) are reduced by 50% versus virus control wells. Times are indicated.</p

Accumulation of recombinant 2G12-KDEL and 89.6P gp140ΔCFI-KDEL env in leaves using the <i>Nb/</i>p19 transient plant expression system.

<p>Extracts produced from the equivalent of 3 mg (2G12-KDEL) (A) and 4.5 mg 89.6Pgp140ΔCFI env (B) of leaf biomass harvested from days 6–24 post-infiltration were loaded onto the gel and proteins detected by anti-KDEL Western blotting.</p

Comparison of the neutralization titers of <i>N.tabacum</i>-derived and CHO-derived mAbs using pseudovirus-based TZM-bl cells.

<p>All leaves were harvested at day 4. IC50s represent the concentration at which relative luminescence units (RLUs) are reduced by 50% versus virus control wells.</p