Human Anti-HIV-1 Env Recombinant Antibody (clone 35O22) (CAT#: PABL-141)

Figure 1 Neutralizing activity of antibodies against a 181-isolate Env-pseudovirus panel. Dendrograms indicate the gp160 protein distance of HIV-1 primary isolate Env glycoproteins. Data below the dendrogram show the number of tested viruses, the percentage of viruses neutralized and the geometric mean or median IC50 for viruses neutralized with an IC50 < 50 μg ml−1.

Huang, J., Kang, B. H., Pancera, M., Lee, J. H., Tong, T., Feng, Y.,... & Doria-Rose, N. A. (2014). Broad and potent HIV-1 neutralization by a human antibody that binds the gp41–gp120 interface. Nature, 515(7525), 138.

Figure 2 Binding specificity of 35O22

a, Neutralization of HIVJRCSF pseudovirus or variants containing the indicated mutations. b, Binding to BG505 SOSIP.664 trimer produced in cells treated with kifunensine or deficient in glycan processing (293S). c, Binding to BG505 trimers with the indicated mutations. BG505 SOSIP.SEKS lacks the furin cleavage site. BG505 WT.664 lacks stabilizing mutations and the antigen primarily represents gp41. d, SPR analysis of binding to immobilized BG505 SOSIP.664 trimers. e, Binding of 35022 (250 nM) to BG505 SOSIP.664 trimers, gp120-gp41ECTO protomers, or monomeric gp120. f, Binding to BG505 SOSIP.664, BG505 SOSIP.SEKS, or the BG505 WT.SEKS lacking the SOSIP mutations.

Huang, J., Kang, B. H., Pancera, M., Lee, J. H., Tong, T., Feng, Y.,... & Doria-Rose, N. A. (2014). Broad and potent HIV-1 neutralization by a human antibody that binds the gp41–gp120 interface. Nature, 515(7525), 138.

Figure 3 Binding or neutralization in the context of a lipid membrane

a, Staining of cell surface expressed HIVJRFL Env. b, ELISA assay of antibody binding to WT or SOS HIVJRFL VLPs. The CD4-inducible 39F antibody or gp41-specific 7B2 are used as controls. c, Access to the HIVJRFL Env trimer on pseudovirions based upon washing the antibody-pseudovirion mixture prior to infecting cells. d, Kinetic assay of HIVJRFL neutralization. See Methods for a description of individual formats. e, Schematic of conformational change resulting in raising of the trimer spike required to permit access of 35O22 to its epitope.

Huang, J., Kang, B. H., Pancera, M., Lee, J. H., Tong, T., Feng, Y.,... & Doria-Rose, N. A. (2014). Broad and potent HIV-1 neutralization by a human antibody that binds the gp41–gp120 interface. Nature, 515(7525), 138.

Figure 4 Analysis of 35O22 autoreactivity

a, Reactivity of 35O22 with HEP-2 epithelial cells. 2F5 was used as a positive control and 17b as a negative control. Antibody concentration was 25 μg/ml. All pictures are shown at 400x magnification. b, SPR analysis of 35O22 binding to anionic phospholipids. 35O22 was injected over PC-CLP liposomes or PC-PS liposomes immobilized on the BIAcore L1 sensor chip. 4E10 and 2F5 were used as positive controls and 13H1 as a negative control. c, Reactivity of 35O22 with autoantigens detected in Luminex assay. 4E10 was used as a positive control. Synagis, an anti-RSV monoclonal antibody, was used as a negative control. SSA, Sjogren's syndrome antigen A; SSB, Sjogren syndrome antigen B; Sm, Smith antigen; RNP, ribonucleoprotein; Scl 70, scleroderma 70; Jo1, antigen; CentrB, centromere B. A positive response is>120 units.

Huang, J., Kang, B. H., Pancera, M., Lee, J. H., Tong, T., Feng, Y.,... & Doria-Rose, N. A. (2014). Broad and potent HIV-1 neutralization by a human antibody that binds the gp41–gp120 interface. Nature, 515(7525), 138.

Figure 5 Effect of sCD4 and sCD4/17b on binding of antibodies 35O22 and PGT151 to BG505 SOSIP.664 by SPR.

The structure of a prefusion mature closed state of HIV-1 provides a critical addition to the pantheon of HIV-1 Env structures with atomic-level detail. Moreover, antibodies 35O22 and PGT151, which bind specifically to the trimeric prefusion conformation of gp41, provide new tools by which to assess the conformational state of gp4113,107,109. The binding of antibodies 35O22 and PGT151 to BG505 SOSIP.664 trimer was tested in the presence of the CD4 receptor and the 17b antibody110 (a co-receptor surrogate which recognizes a bridging sheet epitope that overlaps the site of co-receptor recognition). In the case of antibody 35O22, CD4 binding to the BG505 SOSIP.664 trimer impacted the kinetics, affinity and stoichiometry of binding. 35O22 bound to BG505 SOSIP.664 with an 8.4-fold reduced affinity, primarily contributed by an increased rate of dissociation. The overall binding level (Rmax) normalized to the average level of trimer captured (see also panel d) was lower suggesting substoichiometric binding. Capturing the trimer on a CD4-Ig surface reduced normalized Rmax for PGT151 compared to the 2G12 capture format, suggesting reduced stoichiometry for PGT151 binding to trimer pre-bound with CD4, although kinetics and affinity of interaction were similar. A BG505 SOSIP.664 trimer + sCD4 complex captured onto a 17b surface bound 35O22 but showed no detectable binding to PGT151.

Pancera, M., Zhou, T., Druz, A., Georgiev, I. S., Soto, C., Gorman, J.,... & Stewart-Jones, G. B. (2014). Structure and immune recognition of trimeric pre-fusion HIV-1 Env. Nature,514(7523), 455.