Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (CAT#: TAB-H25)

Figure 1 AGS-22M6 antibody specifically recognizes cell-surface human nectin-4 and its orthologs.

Cell counts of PC-3 cells expressing human, monkey, and rat nectin-4 (open histograms) were overlaid against mocktransfected PC-3 cells (solid histogram).

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer research, 76(10), 3003-3013.

Figure 2 FC measures the binding of AGS-22M6 antibody to human nectin-4.

Saturation curve of AGS-22M6 and AGS-22M6E binding to nectin-4 expressed on T-47D by flow cytometry shows KD of 0.01 nmol/L. MFI, mean fluorescence intensity

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer research, 76(10), 3003-3013.

Figure 3 AGS-22M6E ADC shows potent and specific in vitro cytotoxicity.

PC-3 cells engineered to express human were incubated with increasing concentrations of unconjugated AGS-22M6.

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer research, 76(10), 3003-3013.

Figure 4 T-47D breast cancer cells were killed by AGS-22M6E ADC.

T-47D breast cancer cells, were incubated with increasing concentrations of unconjugated AGS-22M6.

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer research, 76(10), 3003-3013.

Figure 5 Nectin-4–targeted ADC shows significant tumor growth inhibition in multiple xenograft models.

AG-B1 subcutaneous xenografts treated with AGS-22M6E (or controls) at 0.8 mg/kg.

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer researc 76(10), 3003-3013.

Figure 6 AGS-22M6E ADC shows significant tumor growth inhibition in multiple xenograft models.

AG-Br7 orthotopic xenografts treated with AGS-22M6E (or controls) at 5 mg/kg.

Challita-Eid, P. M., Satpayev, D., Yang, P., An, Z., Morrison, K., Shostak, Y., ... & Capo, L. (2016). Enfortumab vedotin antibody–drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer researc 76(10), 3003-3013.

Figure 1 Anti-Human NECTIN4 Therapeutic Antibody (TAB-H25) in DB

Dot Blot analysis of TAB-H25 was performed by coating with human NECTIN4 protein (His tag).

TAB-H25 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG was used as a secondary antibody (1: 8000)

Figure 2 Anti-Human NECTIN4 Therapeutic Antibody (TAB-H25) in WB

Western blot analysis of TAB-H25 was performed with human NECTIN4 protein onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with TAB-H25 and HRP goat anti-Human IgG was used as a secondary antibody (1: 8000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.1 μg)
Lane 2: Reduced antigen (0.3 μg)
Lane 3: Reduced antigen (0.6 μg)

Figure 3 Anti-Human NECTIN4 Therapeutic Antibody (TAB-H25) in ELISA

ELISA analysis of TAB-H25 was performed by coating with human NECTIN4 protein (His tag). Then blocked with BSA and incubated with TAB-H25. The HRP-conjugated goat anti-Human IgG was used as a secondary antibody (1: 5000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 4 Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (TAB-H25, lot#CB1022JF01) in SDS-PAGE

SDS-PAGE analysis of TAB-H25 in reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of TAB-H25 was greater than 95% as determined by SDS-PAGE.

Figure 5 Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (TAB-H25, lot#CB1022JF01) in HPLC

The purity of TAB-H25 was greater than 95% as determined by SEC-HPLC.

Figure 6 Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (TAB-H25, lot#CB1022JF01) in WB

Western blot analysis of TAB-H25 was performed by loading Recombinant NECTIN4 protein.

TAB-H25 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-goat anti-human IgG
Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.3 μg)
Lane 3: Reducing Antigen (0.6 μg)

Figure 7 Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (TAB-H25, lot#CB1022JF01) in ELISA

ELISA analysis of TAB-H25 was performed by coating with Recombinant Human NECTIN4 Protein. Then blocked with BSA and incubated with Human Anti-NECTIN4 Recombinant Antibody (clone Enfortumab) (TAB-H25). The Goat Anti-Human IgG-HRP as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.