Human Anti-HIV-1 Env Recombinant Antibody (clone PGT151) (CAT#: PABL-153)

Figure 1 PGT151–158 neutralization of an indicator panel of 5 pseudoviruses

Serial dilutions of antibody were preincubated with pseudovirus for 1 hour and then added to TZM-bl cells. Three days post-infection luciferase values were measured and % neutralization was calculated.

Falkowska, E., Le, K. M., Ramos, A., Doores, K. J., Lee, J. H., Blattner, C.,... & Mcbride, R. (2014). Broadly neutralizing HIV antibodies define a glycan-dependent epitope on the prefusion conformation of gp41 on cleaved envelope trimers.Immunity, 40(5), 657-668.

Figure 2 PGT151 and PGT152 binding to different forms of Env

(A) PGT151 and PGT152 do not bind to JR-FL gp120 monomer as measured by ELISA. (B) PGT151 binds to native, cleaved (cl) but not to uncleaved (uncl) JR-FL E168K trimers expressed on the surface of 293T cells as measured by flow cytometry, and (C) PGT151 and PGT152 bind to cleaved soluble BG505 SOSIP.664 gp140 trimers but not to the corresponding uncleaved gp140 trimers as measured by ELISA.

Falkowska, E., Le, K. M., Ramos, A., Doores, K. J., Lee, J. H., Blattner, C.,... & Mcbride, R. (2014). Broadly neutralizing HIV antibodies define a glycan-dependent epitope on the prefusion conformation of gp41 on cleaved envelope trimers.Immunity, 40(5), 657-668.

Figure 3 PGT151-PGT158 bind complex carbohydrates

(A) PGT151 and PGT152 neutralization of JR-CSF pseudovirus generated (i) in the presence of the glycosidase inhibitor swainsonine, which prevents the formation of complex glycans by inhibiting the trimming of mannose residues from the Man-α6 arm of the GlcNAcMan5GlcNAc2 structure or (ii) in 293S cells (GNT1−/− cells), a cell line that is deficient in N-acetylglucosaminyltransferase I and is unable to add a GlcNAc residue to the Man5GlcNAc2 structure to permit processing to complex glycans. Neutralization of JR-SCF pseudovirus by 2G12 is not affected by either (i) or (ii) since it binds exclusively to high-mannose glycans on the glycan shield of Env. (B) Glycan microarray analysis reveals PGT151-PGT158 preferentially bind tetraantennary complex carbohydrates and PGT151 additionally binds a triantennary glycan as detected on the Wong glycan array. (C) Glycan microarray analysis using the CFG microarray additionally reveals that all PGT151 family MAbs except PGT153 preferentially bind triantennary complex carbohydrates; and (D) PGT151 and PGT152 bind a tetrantennary complex carbohydrate on the neoglycolipid microarray. The symbols for common monosaccharides are as follows: purple diamonds represent sialic acid, yellow circles represent galactose, red triangles represent fucose, blue squares represent N-acetyl glucosamine and green circles represent mannose.

Falkowska, E., Le, K. M., Ramos, A., Doores, K. J., Lee, J. H., Blattner, C.,... & Mcbride, R. (2014). Broadly neutralizing HIV antibodies define a glycan-dependent epitope on the prefusion conformation of gp41 on cleaved envelope trimers.Immunity, 40(5), 657-668.

Figure 4 PGT151 neutralization of viruses containing N611A, N637A, E647A single and double alanine substitutions

Serial dilutions of antibody (A) PGT151 and (B) PGV04, used as a control, were preincubated with pseudovirus for 1 hour and then added to TZM-bl cells. Three days post-infection luciferase values were measured and % neutralization was calculated.

Falkowska, E., Le, K. M., Ramos, A., Doores, K. J., Lee, J. H., Blattner, C.,... & Mcbride, R. (2014). Broadly neutralizing HIV antibodies define a glycan-dependent epitope on the prefusion conformation of gp41 on cleaved envelope trimers.Immunity, 40(5), 657-668.

Figure 5 ADCC by PGT151 and PGT152

(A) PGT151 and (B) PGT152 were titered for ADCC activity against target cells infected with HIV-1 NL4–3, YU2, JR-CSF and SIVmac239, and using an NK cell line (KHYG-1) expressing CD16 as the effector cell. The killing of virus-infected cells by ADCC is indicated by a loss of relative light units (RLU). SIVmac239 is used as a negative control and HIVIG is used as a positive control.

Falkowska, E., Le, K. M., Ramos, A., Doores, K. J., Lee, J. H., Blattner, C.,... & Mcbride, R. (2014). Broadly neutralizing HIV antibodies define a glycan-dependent epitope on the prefusion conformation of gp41 on cleaved envelope trimers.Immunity, 40(5), 657-668.

Figure 6 B) JRFL gp140 NFLP with 1-3x G4S linkers were screened by IP with selected mAbs, Trimer-specific bNAbs are highlighted in red. (C). IP of BG505 gp140 NFL2P from crude cell supernatant by selected mAbs (trimer-specific mAbs highlighted in red). Note that the IP band intensity for non-NAb F105 is relatively light compared to those of the bNAbs, suggesting the majority of expressed protein is well-ordered trimers.

Sharma, S. K., de Val, N., Bale, S., Guenaga, J., Tran, K., Feng, Y.,... & Wyatt, R. T. (2015). Cleavage-independent HIV-1 Env trimers engineered as soluble native spike mimetics for vaccine design. Cell reports, 11(4), 539-550.