Anti-Human HIV V3 Recombinant Antibody (KD-247) (CAT#: TAB-357CL)

Figure 1 Selection of neutralization-resistant virus variants against KD-247.

Sensitivity of MOKW5p (200) and MOKW9p (2000) virus to KD-247 as determined by MTT assay. PM1/CCR5 cells (2 × 10³) were exposed to 100 TCID50 of MOKW9p(-), MOKW5p(200), or MOKW9p(2000) virus and were cultured in the presence of various concentrations of KD-247. The IC50 values were determined by MTT assay on day 7 of culture. The assay was conducted in duplicate. The values shown are representative of three separate experiments.

Shibata, J., Yoshimura, K., Honda, A., Koito, A., Murakami, T., & Matsushita, S. (2007). Impact of V2 mutations on escape from a potent neutralizing anti-V3 monoclonal antibody during in vitro selection of a primary human immunodeficiency virus type 1 isolate. Journal of virology, 81(8), 3757-3768.

Figure 2 Neutralization sensitivities of pseudoviruses with env genes from passaged MOKW viruses to MAbs, rsCD4, and CCR5 inhibitors.

KD-247 was preincubated with 100 TCID50 of each MOKW pseudotype virus for 15 min, followed by the addition of the mixtures to the target cells (GHOST-hi5). The inhibitory effects were determined by measuring the luciferase activities on day 2 of culture. Conc, concentration.

Shibata, J., Yoshimura, K., Honda, A., Koito, A., Murakami, T., & Matsushita, S. (2007). Impact of V2 mutations on escape from a potent neutralizing anti-V3 monoclonal antibody during in vitro selection of a primary human immunodeficiency virus type 1 isolate. Journal of virology, 81(8), 3757-3768.

Figure 3 Comparison of antibody binding to cell surface-expressed MOKW Env proteins.

293T cells transfected with MOKW Env expression vectors were harvested at 24 h posttransfection and stained with KD-247. Flow cytometry data for binding of the KD-247 (black lines) to cell surface MOKW Env proteins are shown among GFP gated 293T cells.

Shibata, J., Yoshimura, K., Honda, A., Koito, A., Murakami, T., & Matsushita, S. (2007). Impact of V2 mutations on escape from a potent neutralizing anti-V3 monoclonal antibody during in vitro selection of a primary human immunodeficiency virus type 1 isolate. Journal of virology, 81(8), 3757-3768.

Figure 4 Binding affinity of anti-V3 MAbs to monomeric gp120.

Viral lysates for each MOKW pseudovirus were used. gp120 was captured onto microtiter wells using a sheep polyclonal antibody specific for the C terminus of gp120. Serial dilutions of KD-247 was tested for binding by ELISA. Because of differences in the amount of bound gp120, optical density at 405 nm (OD405) values were normalized to saturating levels of antibody (5 μg/ml) for comparison. Conc, concentration.

Shibata, J., Yoshimura, K., Honda, A., Koito, A., Murakami, T., & Matsushita, S. (2007). Impact of V2 mutations on escape from a potent neutralizing anti-V3 monoclonal antibody during in vitro selection of a primary human immunodeficiency virus type 1 isolate. Journal of virology, 81(8), 3757-3768.

Figure 5 The gp120 mutation profile of KD-247 HIV-1BaL evasion variants for 14 additional passages without KD-247.

The ratioof the PNGS insertion in the V2 region and mutations in the C2 and V3 region of HIV-1BaL evasion variants in gp120 were plotted for 16 passages in the presence of KD-247 and an additional 14 passages in the absence of the mAb. The y-axis showsthe percentage of PNGS insertions or mutations in the tested clones. The x-axis shows the concentration of KD-247.

Hatada, M., Yoshimura, K., Harada, S., Kawanami, Y., Shibata, J., & Matsushita, S. (2010). Human immunodeficiency virus type 1 evasion of a neutralizing anti-V3 antibody involves acquisition of a potential glycosylation site in V2. Journal of General Virology, 91(5), 1335-1345.