NeuroMab™ Anti-CXCR4 BBB Shuttle Antibody(NRZP-1022-ZP3796)

Figure 1 shows the binding of human anti-CXCR4 antibodies F7, F9, D1 and E2 to CEM cells expressing native human CXCR4 on the cell surface.

Figure 2 shows antibody competition for binding to CEM cells between FITC-labeled anti-CXCR4 antibody F9 and a panel of unlabeled human anti-CXCR4 antibodies.

Figure 3 shows flow cytometry analysis of BMS-936564 binding.

Figure 4 shows that anti-CXCR4 human antibodies F7 (BMS-936564), F9 and D1 inhibit the binding of125I-labeled CXCL12 to CXCR4 expressed on CEM cells. E2 antibody does not inhibit the binding of CXCL12 to CEM cells.

Figure 5 shows the inhibition of CXCL12(SDF-1) induced calcium flux in CEM cells by anti-CXCR4 human antibodies F7 (BMS-936564), F9 and D1. E2 did not significantly inhibit CXCL12-induced calcium flux.

Figure 6 shows inhibition of CXCL12-induced calcium flux in CXCR4+ cells by anti-CXCR4 antibody MDX-1338 (BMS-936564) or anti-CXCL12 antibody.

Calcium flux assays were performed by incubating Ramos cells with calcium 4 dye in the presence or absence of test antibodies or isotype controls. Dye-loaded cells were incubated with Ramos and CEM cells at room temperature with 50 nM and 5 nM CXCL12, respectively. The area under the fluorescence curve between 20 and 200 seconds was quantified and the EC50 was calculated.

Figure 7 shows the inhibition of CXCL12-induced migration of CEM cells by anti-CXCR4 human antibodies F7 (BMS-936564) and F9, whereas antibodies D1 and E2 did not significantly inhibit migration.

Figure 8 shows the in vitro inhibition of Ramos tumor cell proliferation by anti-CXCR4 human antibodies F7 (BMS-936564), F9 and E2.

Figure 9 shows the inhibition of Ramos tumor cell proliferation in vivo in a subcutaneous tumor model by anti-CXCR4 human antibodies F7 (BMS-936564) and F9.

Figure 10 shows the percentage survival of mice treated with anti-CXCR4 human antibody F9 or anti-CXCR4 antibody BMS-936564 in the Ramos systemic tumor cell model.

BMS-936564 was very effective in this Ramos systemic model, whereas anti-CXCL12 Ab showed no effect.