NeuroMab™ Anti-ADAM17 BBB Shuttle Antibody, Clone D8P1C1

Figure 1. Results of experiments performed with the human breast cancer cell line MDA-MB-231.

Fig. A shows the results of experiments in which the effects of various anti-ADAM17 antibodies or antibody fragments on viability of MDA-MB-231 breast cancer cells were tested using an ALAMARBLUE cell viability assay. The effects of D5 IgG, D5 Fab, D8 IgG, and D8 Fab were tested. The control was an IgGl control. GM6001is a matrix metalloprotease inhibitor.

Figure 2. Experimental results using the human breast cancer cell line MDA-MB-231.

Fig. B-D present the results of experiments in which cultured MDA-MB-231 cells were stained with anti-ADAM17 antibodies D5 and D8 and alexa568-labeled anti-human IgG secondary antibody (green). As a control, cells were stained with the secondary antibody only. Nuclei were stained with Hoechst (blue) before imaging by confocal microscopy. Microscopic images of MDA-MB-231 cells stained for D5 (Figure 2B), D8 (Figure 2C), and control (Figure 2D) are shown, respectively.

Figure 3 shows the results of a dose-response experiment performed to assess the effect of the parental D8 clone and the three most potent affinity-matured clones derived from D8 on the proliferation of the MDA-MB-231 cell line - Quantitative Cell Viability Assay Using ALAMARBLUE .

Figure 4 shows the results of a dose-response experiment performed to assess the effect of two different batches of parental D8 clones (referred to as "D8 (NIH)" and "D8 (LP)" in the figure), and two Effect of the most potent affinity matured clones of D8 (ie, "D8.PI.Cl" and "D8.P2.C6.") on the proliferation of the MDA-MB-231 cell line - quantified using the ALAMARBLUE cell viability assay.

Figure 5. Proliferation inhibition of ovarian cancer cell line SKOV3 by anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1C1.

Figure 6. Proliferation inhibition of ovarian cancer cell line CaoV3 by anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1Cl.

Figure 7. Proliferation inhibition of the ovarian cancer cell line OVCAR-3 by the anti-ADAM17 monoclonal antibody D8 and its affinity-matured version, D8P1C1.

Figure 8. Proliferation inhibition of breast cancer cell line SKBR-3 by anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1C1.

Figure 9. Proliferation inhibition of breast cancer cell line MCF-7 by anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1C1.

Figure 10. Proliferation inhibition of colon cancer cell line LIM1215 by anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1C1.

Figure 11. Proliferation inhibition of the glioblastoma cell line U87 MG by the anti-ADAM17 monoclonal antibody D8 and its affinity matured version D8P1C1.

Figure 13. Data showing the antitumor effect of mAh D8P1C1 in a triple negative breast cancer xenograft assay.

Inject 10 million MDA-MB-231 cells per mouse into 6-8 week old athymic nude mice (n=5). In the treatment group ("D8" in the figure), each mouse was injected with D8P1C1 (i.p.) at a dose of 15 mg/kg every two weeks for 4 weeks. In the control group, PBS was used as vehicle control ("Control" in the figure). The average tumor volume of the control and treatment groups was measured. The graph shows mean tumor volume (mm3) plotted against days post-injection. On the last day of the study, the percent (%) inhibition of tumor growth by D8P1C1 was 76.8% (calculated as 100 c [1 - [(TreatedFinal day - TreatedDay 1)/(ControlFinal day - ControlDay 1)]]).