NeuroMab™ Anti-NOTCH3 Antibody(NRP-0422-P1859)

Figure 1: Affinity-matured mAbs that bind the ligand-binding domain of Notch1 inhibit the binding of DLL4 to Notch1.

FACS analysis demonstrated that various members of a series of affinity matured mAbs that bind the ligand binding domain of Notch1 potently block binding between DLL4 and Notch1.

Figure 2: Identification of fAbs that bind the ligand-binding domain of Notch3 and effectively block Notch ligand binding.

FACS analysis demonstrated that fAbs binding to the ligand-binding domain of Notch3 identified by phage display were able to block the binding of the Notch ligand (JAG1) to HEK 293 cells co-transfected with GFP and Notch3.

Figure 3: Monoclonal antibodies to the ligand-binding domain of Notch inhibit Notch signaling.

Monoclonal antibodies against the Notch ligand binding domain inhibit Notch signaling in response to Notch ligand DLL4. Western blot analysis of Notch ICD formation as indicative of Notch signaling confirmed that the presence of DLL4 induced robust formation of ICD (compare −DLL4 and +DLL4), and addition of a gamma secretase inhibitor (DBZ) blocked this signaling event. Antibodies to the ligand binding region of Notch (90R21, 90R22, and R9029) also inhibited ICD formation. In contrast, antibodies that bind Notch outside the ligand biding epitope (13M57) and non-binding antibodies (control mAb) have little or no affect on ICD formation.

Figure 4: Monoclonal antibodies to the ligand-binding domain of Notch inhibit tumor growth in vivo.

NOD/SCID mice were injected with dissociated PE13 breast tumor cells and treated with either control antibodies (black squares); anti-Notch ligand binding region antibodies 90R21 (open circles); control antibodies in combination with Paclitaxel (black triangles); or 90R21 antibodies in combination with Paclitaxel (open triangles). Treatment with 90R21 antibodies reduced tumor growth significantly as compared to control antibody treated animals, while the combination with Paclitaxel virtually eliminated tumor growth.

Figure 5: Monoclonal antibodies to the ligand-binding domain of Notch inhibit tumor growth in vivo.

NOD/SCID mice were injected with dissociated T3 breast tumor cells and treated with either control antibodies (black squares); anti-Notch ligand binding region antibodies 90R21 (open circles); control antibodies in combination with Paclitaxel (black triangles); or 90R21 antibodies in combination with Paclitaxel (open triangles). Treatment with 90R21 antibodies reduced tumor growth significantly as compared to control antibody treated animals, while the combination with Paclitaxel virtually eliminated tumor growth.