NeuroMab™ Anti-Notch 1 BBB Shuttle Antibody(NRZP-1022-ZP3248)

Figure 1: Binding of anti-NOTCH1 monoclonal antibodies 13M57 and 31M80 to cell surface expressed NOTCH1.

FACS analysis of cells co-expressing full length NOTCH1 receptor and GFP incubated with, from left to right, IgG1 control antibodies, 13M57 and anti-human NOTCH1 EGF 1-5 antisera.

Figure 2: Binding of anti-NOTCH1 monoclonal antibodies 13M57 and 31M80 to cell surface expressed NOTCH1.

Anti-NOTCH1 antibodies 13M57 and 31M80 showed increased binding to PE13 breast tumor cells compared to an isotype control antibody (bottom) and this binding corresponded to cells that express high levels of ESA and CD44 (top: CD44 x-axis and NOTCH1 y-axis).

Figure 3: Antibodies against NOTCH1 EGF13 and EGF4 do not effectively block ligand binding.

NOTCH1 expressing HEK 293 cells were incubated with either DLL4-Fc (left) or JAG1-Fc (right) in the presence of anti-NOTCH1 antibodies (13M57, 31M103, 31M106, or 31M108) or control anti-DLL4 (21M18) or anti-JAG1 (64M14) antibodies.

Figure 4: Effect of NOTCH1 mAb against EGF4 on breast tumor cell growth in vitro.

Breast tumor cells were cultured for three days in the presence of 2.5 ug/mL or 5 ug/mL anti-NOTCH1 antibody, control mouse IgG, or no antibody, followed by 18 hr BrdU labeling.

Figure 5: Effect of NOTCH1 monoclonal antibody 13M57 on PE-13 tumor cells in vivo.

NOD/SCID mice injected with PE-13 tumor cells were treated with PBS or anti-NOTCH1 antibodies 3 days after cell injection, and the growth of tumor cells was determine twice a week.

Figure 6: Effect of NOTCH1 monoclonal antibody on colon tumor cells in vivo.

NOD/SCID mice injected with OMP-C9 colon tumor cells were treated with PBS or anti-NOTCH1 13M57 antibodies three days after cell injection, and the growth of tumor cells was determine twice a week.

Figure 7: Effect of NOTCH1 monoclonal antibody 13M57 on luciferase-expressing PE-13 tumor cells.

Quantification of luciferase signal shows total tumor volume was significantly reduced (p=0.04) in animals treated with anti-NOTCH1 antibodies 13M57 compared to PBS and 5M108 injected controls.

Figure 8: Effect of anti-NOTCH1 antibody 13M57 and chemotherapy combination therapy on breast tumor recurrence.

Figure 9: Effect of combination therapy with anti-NOTCH1 antibody 13M57 and chemotherapy on colon tumor growth.

Figure 10: Effect of combination therapy with anti-NOTCH1 antibody 13M57 and chemotherapy on growth of established colon tumors.