Chimeric (Mouse/Human) Anti-TNFRSF8 Recombinant Antibody (TAB-153) (CAT#: TAB-153)

Figure 6 Brentuximab vedotin reduces viable cell number in CD30-positive but not CD30-negative GCT cells.

The CD30-positive EC cell lines GCT27 and NCCIT as well as the CD30-negative choriocarcinoma line JAR were treated with MMAE (A) or brentuximab vedotin (B-D). GCT27 (B), JAR (C) and NCCIT (D) were exposed to 250 ng/ml brentuximab vedotin. After 24, 48, 72 and 96 hrs of culture, cells were resuspended in equal volume for analysis. Viable Hoechst-negative cells were enumerated for 180 sec. by flow cytometry and are represented as multiples (x-fold) of the untreated control obtained at 24 hrs. To further evaluate dose-dependent effects to brentuximab vedotin, the three cell lines were exposed for 96 hrs to 250, 500 and 1000 ng/ml of the ADC as well as 100 pM MMAE (E). Enumerated viable Hoechst-negative cells are expressed in percent of the untreated control at 96 hrs.

Götz, B., van Beekum, C., Nettersheim, D., Schorle, H., Calaminus, G., Leuschner, I.,... & Schönberger, S. (2015). brentuximab Vedotin Presents Profound Anti-tumor Efficacy In Cd30+ And Co-cultured Cd30-Germ Tumor Cells: o-129. Pediatric Blood & Cancer, 62, S179.

Figure 7 Brentuximab vedotin exerts pronounced bystander activity on MMAE-sensitive, CD30-negative GCT cells in coculture with CD30-positive embryonal carcinoma.

For determination of bystander efficacy after drug exposure, cells were stained with CSFE and anti-CD30.PE (BER-H2, eBiosience/Germany). After 96 hrs of drug exposure, cell cultures were resuspended in equal volume and acquired for 180 sec. by flow cytometry. CD30-negative JAR cells are EPCAM positive (y-axis) but CD30 negative (x-axis) while GCT27 cells are EPCAM and CD30 positive. Hoechst-negative viable CD30-positive and CD30-negative subpopulations were assessed separately after gating on the respective cell fraction (A). Viable cell numbers are expressed in per cent of untreated control (B). Proliferation is investigated by CSFE dilution upon cellular division. To indicate inhibition of proliferation MFI of experimental conditions was normalized to the MFI of untreated cells and presented as x-fold MFI (C). Cell death was quantified as the proportion of Hoechstpositive cells of the entirety of acquired cells (D)

Götz, B., van Beekum, C., Nettersheim, D., Schorle, H., Calaminus, G., Leuschner, I.,... & Schönberger, S. (2015). brentuximab Vedotin Presents Profound Anti-tumor Efficacy In Cd30+ And Co-cultured Cd30-Germ Tumor Cells: o-129. Pediatric Blood & Cancer, 62, S179.

Figure 1 Anti-Human CD30 Therapeutic Antibody (TAB-153) in ELISA

ELISA analysis of TAB-153 was performed by coating with Recombinant Human CD30 Protein. Then blocked with BSA, and incubated with Anti-Human CD30 Antibody (TAB-153). The HRP-conjugated goat anti-human IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 2 Chimeric (mouse/Human) Anti-TNFRSF8 Recombinant Antibody (TAB-153) in ELISA

ELISA analysis of TAB-153 was performed by coating with CD30, Human, Recombinant.
The secondary antibody: HRP-Anti-Human IgG (H+L)