Anti-Human NOTCH1 Recombinant Antibody (23814) (CAT#: TAB-545MZ)

Figure 1 In vitro characterization of 23814 activity.

Notch1 luciferase reporter line cocultured with stable lines expressing Jag1, Jag2, DLL1, and DLL4 ligands in the presence of 23814 or control hIgG antibodies. Results represented as percent inhibition of luciferase activity.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 2 In vitro characterization of 23814 activity.

Karpas 45 plated on wells coated with recombinant Jag1, Jag2, DLL4, or Fc (no ligand) in the presence of 23814 or hIgG control (10 μg/mL). Lane 1, no ligand + hIgG; lane 2, ligand + hIgG; lane 3, ligand + 23814 antibody. Western blots were probed with antibody specific for cleaved Notch1 ICD (Cell Signaling Technology). Equal amounts of lysate transferred to a separate blot were probed with β-tubulin antibody as a loading control.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 3 In vitro characterization of 23814 activity.

Karpas 45 cocultured with ligand-expressing CHO lines in the presence of 23814 or hIgG.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 4 In vitro characterization of 23814 activity.

FACS analysis of 23814 binding to CHO cells expressing human or murine Notch1, human Notch2, or human Notch3.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 5 23814 inhibits Notch1 signaling in vivo and does not cause gut toxicity.

23814 treatment of mice (n = 5 animals/group) results in thymocyte depletion.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 6 23814 inhibits Notch1 signaling in vivo and does not cause gut toxicity.

23814 effectively inhibits Notch1 signaling in vivo, no weight loss occurs.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 7 23814 inhibits growth of a renal clear cell PDX tumor.

NCR nude mice (n = 10) received either hIgG control, anti-ErbB3 (AV-203), 23814, or tivozanib. 23814 and tivozanib monotherapy significantly inhibited tumor growth relative to control.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 8 23814 treatment increases tumor vasculature.

the HT-1080 tumor model (n = 10) was treated with control hIgG, 23814, tivozanib, or a combination of 23814 plus tivozanib. Combination treatment results in significantly greater tumor growth inhibition than either tivozanib or 23814 alone.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 9 Combined inhibition of Notch1 and VEGFR pathways enhances antitumor efficacy in models partially resistant to VEGFR inhibitor monotherapy.

BH224 tumor models (n = 10) exhibit a range of responses to treatment with 23814, tivozanib, or a combination of the two.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 10 Combined inhibition of Notch1 and VEGFR pathways enhances antitumor efficacy in models partially resistant to VEGFR inhibitor monotherapy.

BH226 tumor models (n = 10) exhibit a range of responses to treatment with 23814, tivozanib, or a combination of the two.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 11 Combined inhibition of Notch1 and VEGFR pathways enhances antitumor efficacy in models partially resistant to VEGFR inhibitor monotherapy.

BH413 tumor models (n = 10) exhibit a range of responses to treatment with 23814, tivozanib, or a combination of the two.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 12 Unsupervised hierarchical clustering identifies tumors likely to respond to 23814/tivozanib combination therapy.

BH228 tumor growth inhibition is significantly increased by combination treatment with 23814/tivozanib relative to tivozanib monotherapy.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 13 Unsupervised hierarchical clustering identifies tumors likely to respond to 23814/tivozanib combination therapy.

BH270 tumor growth inhibition is significantly increased by combination treatment with 23814/tivozanib relative to tivozanib monotherapy.

Proia, T., Jiang, F., Bell, A., Nicoletti, R., Kong, L., Kreuter, K., ... & Perino, S. (2015). 23814, an inhibitory antibody of ligand-mediated Notch1 activation, modulates angiogenesis and inhibits tumor growth without gastrointestinal toxicity. Molecular cancer therapeutics, 14(8), 1858-1867.

Figure 1 Anti-Human NOTCH1 Antibody (TAB-545MZ) in SDS-PAGE

Lane 1: Non-reduced mAb
Lane 2: Reduced mAb

Figure 2 Anti-Human NOTCH1 Antibody (TAB-545MZ) in HPLC

The purity of TAB-545MZ was greater than 95% as determined by SEC-HPLC.