NeuroMab™ Anti-ADAM17 BBB Shuttle Antibody(NRZP-1022-ZP3629)

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 D5 clone and the three most potent affinity-matured clones derived from D5 (i.e. "D5.PI.A4", "D5.P2.All1") . " and "D5.P2.B3") on the proliferation of the MDA-MB-231 cell line - quantified using the ALAMARBLUE cell viability assay.

Figure 4 shows the results of experiments performed to assess the effect of certain antibodies on the proliferation of the ovarian cancer cell line SKOV3 (transduced with the MUC 16 ectodomain), as quantified using the ALAMARBLUE cell viability assay.

Figure 5. Proliferation inhibition of ovarian cancer cell line SKOV3 by anti-ADAM17 mAb D5 and its affinity matured version.

Figure 6. Proliferation inhibition of the ovarian cancer cell line CaoV3 by anti-ADAM17 mAb D5 and its affinity matured version.

Figure 7. Proliferation inhibition of ovarian cancer cell line OVCAR-3 by anti-ADAM17 mAb D5 and its affinity matured version D5P2A11.

Figure 8. Proliferation inhibition of breast cancer cell line SKBR-3 by anti-ADAM17 monoclonal antibody D5 and its affinity matured version.

Figure 9. Proliferation inhibition of breast cancer cell line MCF-7 by anti-ADAM17 mAb D5 and its affinity matured version D5P2A11.

Figure 10. Proliferation inhibition of colon cancer cell line LIM1215 by anti-ADAM17 monoclonal antibody D5 and its affinity matured version.

Figure 11. Proliferation inhibition of the glioblastoma cell line U87 MG by the anti-ADAM17 monoclonal antibody D5 and its affinity matured version.

Figure 12. Data from a FRET-based peptide cleavage assay.

An ADAM17 peptide substrate derived from TNF-α was used. The excitation wavelength is 320nm and the emission wavelength is 405nm. The buffer contains 20 mM Tris pH 8.8, 2 mM zinc chloride, and 50 mM peptide substrate. Data in the graph are the mean of triplicate determinations. The maximum deviation is within 10% of the mean.