Anti-Human CD27 Recombinant Antibody (Varlilumab) (CAT#: TAB-H74)

Figure 1 Varlilumab pharmacokinetics and receptor occupancy.

Serum concentrations of varlilumab were determined by ELISA by using recombinant CD27-coated plates and anti-human IgG-horseradish peroxidase conjugate as a detection probe. Values represent the mean for patients in each cohort.

Burris, H. A., Infante, J. R., Ansell, S. M., Nemunaitis, J. J., Weiss, G. R., Villalobos, V. M., ... & Hawthorne, T. R. (2017). Safety and activity of varlilumab, a novel and first-in-class agonist anti-CD27 antibody, in patients with advanced solid tumors. Journal of Clinical Oncology, 35(18), 2028-2036.

Figure 2 Varlilumab pharmacokinetics and receptor occupancy.

Percent receptor occupancy was determined by flow cytometry using anti-human IgG conjugate for detection of varlilumab and a noncompeting anti–CD27-labeled monoclonal antibody for identifying CD27 expression. Values respresent the percentage of CD8+/CD27+ T cells that stained positive with anti-human IgG treated at different dose levels.

Burris, H. A., Infante, J. R., Ansell, S. M., Nemunaitis, J. J., Weiss, G. R., Villalobos, V. M., ... & Hawthorne, T. R. (2017). Safety and activity of varlilumab, a novel and first-in-class agonist anti-CD27 antibody, in patients with advanced solid tumors. Journal of Clinical Oncology, 35(18), 2028-2036.

Figure 3 Varlilumab pharmacokinetics and receptor occupancy.

Mean level of soluble CD27 as using a sandwich ELISA that is not obstructed by the presence of bound varlilumab. Black triangles (:) represent varlilumab dosing. ELISA, enzymelinked immunosorbent assay; Ig, immunoglobulin; PBMC, peripheral blood mononuclear cell; TIL, tumor-infiltrating lymphocytes.

Burris, H. A., Infante, J. R., Ansell, S. M., Nemunaitis, J. J., Weiss, G. R., Villalobos, V. M., ... & Hawthorne, T. R. (2017). Safety and activity of varlilumab, a novel and first-in-class agonist anti-CD27 antibody, in patients with advanced solid tumors. Journal of Clinical Oncology, 35(18), 2028-2036.

Figure 4 Increase in responses to melanoma antigen in select patients who were treated with varlilumab.

Enumeration of CD8+ T-cell responses in HLA-A2– or HLAA3–positive patients with melanoma to the indicated melanoma-associated antigens. Peripheral blood mononuclear cells (PBMCs) from days 1, 29, and 85 were assayed for interferon-gamma production in response to antigen-presenting cells that were pulsed with selected melanoma antigen peptides after 2 weeks in vitro stimulation with a peptide cocktail. Values shown are the fold change on the basis of dividing the absolute counts for each peptide by the counts with control HIV gag–derived peptides. P values were calculated by using one-way analysis of variance for individual timepoints or unpaired Student t tests across timepoints. For patient 04-9002, PBMCs were directly stained for major histocompatibility complex multimer binding and representative dot plots are presented. ns, not significant.

Burris, H. A., Infante, J. R., Ansell, S. M., Nemunaitis, J. J., Weiss, G. R., Villalobos, V. M., ... & Hawthorne, T. R. (2017). Safety and activity of varlilumab, a novel and first-in-class agonist anti-CD27 antibody, in patients with advanced solid tumors. Journal of Clinical Oncology, 35(18), 2028-2036.

Figure 5 Induced proliferations of T cell subsets and coexpression of costimulatory and coinhibitory receptors (immune checkpoint molecules).

Differences in short term (3 day) or long term (7 day) T cell cultures undergoing stimulations with varlilumab or control antibody (hIgG) show more CD8+ cells (a) entering the division cycle in the varlilumab-treated group. Numbers in the histograms refer to the percent of indicated cells (CD8+ or CD3+CD8-) that are dividing as reflected by CFSE staining (a bottom panel).

Ramakrishna, V., Sundarapandiyan, K., Zhao, B., Bylesjo, M., Marsh, H. C., & Keler, T. (2015). Characterization of the human T cell response to in vitro CD27 costimulation with varlilumab. Journal for immunotherapy of cancer, 3(1), 37.

Figure 6 Induced proliferations of T cell subsets and coexpression of costimulatory and coinhibitory receptors (immune checkpoint molecules).

CD27+ proliferating CD4+ and CD8+ T cells were stained with antibodies to additional cell surface markers representing costimulatory (GITR, OX40, ICOS and 4-1BB) and coinhibitory (PD-1) molecules. Dividing CD8+ and CD4+ T cells show marked upregulation of costimulatory and coinhibitory molecules relative to non-dividing cells. Shaded histograms represent T cells stimulated with control antibody and anti-CD3 (OKT3). Numbers in the histograms refer to percent of T cells (dividing or non-dividing CD8 or CD4 cells) that are positive for the indicated costimulatory or coinhibitory marker.

Ramakrishna, V., Sundarapandiyan, K., Zhao, B., Bylesjo, M., Marsh, H. C., & Keler, T. (2015). Characterization of the human T cell response to in vitro CD27 costimulation with varlilumab. Journal for immunotherapy of cancer, 3(1), 37.

Figure 7 Early detection of cytokines by varlilumab costimulated T cell subsets.

T cells were preactivated with OKT3 for 46 h and post activated for 4 h with OKT3+ varlilumab or OKT3+ hIgG. Intracellular staining for IFNγ response in different T cell subpopulations (a) Naïve CD4+ (CD45RO ̶) and memory (CD45RO+) and (b) naïve CD8+ (CD45RO ̶) and memory (CD45RO+) T cells. Cells were co-stained for the activation marker CD69. Numbers in individual plots refer to percent IFNγ+ cells.

Ramakrishna, V., Sundarapandiyan, K., Zhao, B., Bylesjo, M., Marsh, H. C., & Keler, T. (2015). Characterization of the human T cell response to in vitro CD27 costimulation with varlilumab. Journal for immunotherapy of cancer, 3(1), 37.

Figure 1 Human CD27 Recombinant Antibody (TAB-H74) in SDS-PAGE

SDS-PAGE analysis of TAB-H74 in non-reduced (lane 1) and β-mercaptoethanol-reduced (lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 130-180 kDa.

Figure 2 Human CD27 Recombinant Antibody (TAB-H74) in HPLC

The purity of TAB-H74 was greater than 95% as determined by SEC-HPLC.