Mouse Anti-MS4A1 Recombinant Antibody (clone GA101) (CAT#: PABL-423)

Figure 1 Activities of RTX and GA101 in whole blood assays.

RTX or GA101 were added to B-CLL/B-NHL whole blood samples at 100 μg/ml and percentage cell death of neoplastic B cell targets measured by FACS analysis after 4 (black bars) and 24 h (gray bars).

Bologna, L., Gotti, E., Manganini, M., Rambaldi, A., Intermesoli, T., Introna, M., & Golay, J. (2011). Mechanism of action of type II, glycoengineered, anti-CD20 monoclonal antibody GA101 in B-chronic lymphocytic leukemia whole blood assays in comparison with rituximab and alemtuzumab. The Journal of Immunology, 186(6), 3762-3769.

Figure 2 Activities of RTX and GA101 in whole blood assays.

Percentages of cell death with 100 μg/ml GA101 at 24 h were plotted against CD20 expression levels, measured as number of CD20 molecules/cell, and r² calculated.

Bologna, L., Gotti, E., Manganini, M., Rambaldi, A., Intermesoli, T., Introna, M., & Golay, J. (2011). Mechanism of action of type II, glycoengineered, anti-CD20 monoclonal antibody GA101 in B-chronic lymphocytic leukemia whole blood assays in comparison with rituximab and alemtuzumab. The Journal of Immunology, 186(6), 3762-3769.

Figure 3 GA101 but not RTX induces CD107a on NK cells in normal and B-CLL whole blood.

MNC from five normal donors were incubated for 3 h with 10 μg/ml GA101, RTX, or TX and percentage of CD107a expression on CD56+ NK cells analyzed by flow cytometry.

Bologna, L., Gotti, E., Manganini, M., Rambaldi, A., Intermesoli, T., Introna, M., & Golay, J. (2011). Mechanism of action of type II, glycoengineered, anti-CD20 monoclonal antibody GA101 in B-chronic lymphocytic leukemia whole blood assays in comparison with rituximab and alemtuzumab. The Journal of Immunology, 186(6), 3762-3769.

Figure 4 Ramos cells werestained for 30 minutes at 37°C with Cy-3–labeled GA101 (red fluorescence), which binds to the membraneganglioside GMI in lipid rafts.

CD20 molecules do not redistribute tolipid rafts on binding of GA101 and do not colocalizewith CTB-Alexa 488.

Mössner, E., Brünker, P., Moser, S., Püntener, U., Schmidt, C., Herter, S., ... & Ferrara, C. (2010). Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell–mediated B-cell cytotoxicity. Blood, The Journal of the American Society of Hematology, 115(22), 4393-4402.

Figure 5 GA101 does not mobilize CD20 intolipid rafts.

On binding of GA101 to CD20 in Ramos cells,CD20 is mainly found in the Triton X-100-solublefraction (S) and not in the Triton X-100-insoluble pellet(P) representing lipid rafts (top row). In contrast, bindingof rituximab resulted in the distribution of CD20 into theTriton X-100-insoluble pellet fraction (S; top row). The distribution of Lyn as a typical lipid raft marker and CD71 as a nonlipid raft marker is not affected (bottom2 rows).

Mössner, E., Brünker, P., Moser, S., Püntener, U., Schmidt, C., Herter, S., ... & Ferrara, C. (2010). Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell–mediated B-cell cytotoxicity. Blood, The Journal of the American Society of Hematology, 115(22), 4393-4402.

Figure 6 CDC assays (LDH release) with Z138 mantle cell lymphoma cells.

In the presence of physiologic concentrations of human unspecific IgG (10 mg/mL RedImmune), the type II anti-CD20 antibody GA101 (black) mediates greatly reduced CDC induction compared with rituximab (gray; n=3).

Mössner, E., Brünker, P., Moser, S., Püntener, U., Schmidt, C., Herter, S., ... & Ferrara, C. (2010). Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell–mediated B-cell cytotoxicity. Blood, The Journal of the American Society of Hematology, 115(22), 4393-4402.

Figure 7 Superior antitumor efficacy of GA101 compared with rituximab in human lymphoma xenograft models.

Treatment of the aggressive orthotopic disseminated Z138 (MCL) model was initiated 29 days after intravenous injection of tumor cells (n=10 per group). Treatment with 10 mg/kg GA101 (every 7 days, 6 times, intravenously; black line) resulted in increased overall andmedian survival, compared with 10 mg/kg rituximab treatment (dark gray line; P< .008) and vehicle control (light gray line). šindicates the treatment time points.

Mössner, E., Brünker, P., Moser, S., Püntener, U., Schmidt, C., Herter, S., ... & Ferrara, C. (2010). Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell–mediated B-cell cytotoxicity. Blood, The Journal of the American Society of Hematology, 115(22), 4393-4402.