Afuco™ Anti-Human AGS-16 ADCC Recombinant Antibody (AGS-16M18), ADCC Enhanced (CAT#: AFC-026CL)

Figure 1 In vitro and in vivo effects of anti-ENPP3 ADC.

A, KU812 cells were incubated for 6 days with increasing concentrations of anti-ENPP3 or isotype control Ab conjugated to vcMMAE or mcMMAF,and cell viability was determined. The inset table shows drug-antibody ratios(DAR)and IC50 values for each ADC.B, mice bearing 100 to 150 mm3 UG-K3 RCC xenografted tumors were treated with AGS16-7.8 or AGS16-9.69 conjugated to vcMMAE or mcMMAF at 3 mg/kg by i.v. injection on days 0, 3, 7, and 10.Data show mean ± SEM tumor volume for each group.The inset figure shows a representative image of ENPP3 expression by IHC in a UG-K3 tumor section.

Doñate, F.,Raitano, A., Morrison, K., An, Z., Capo, L., Aviña, H., & Moriya, R.(2016).AGS16F is a novel antibody drug conjugate directed against ENPP3 for the treatment of renal cell carcinoma.Clinical Cancer Research,22(8),1989-1999.

Figure 2 Internalization of AGS16F by KU812 cells.

AGS16F was incubated with KU812 cells,then fixed and stained as indicated. Samples were imaged using laser scanning confocal fluorescence microscopy: AGS16F appears in green;LAMP-1(a marker of lysosomes) appears in red;and the nuclei are shown in blue in the 3-channel Z-stack overlay in the last column.Z-stack overlay images show single plane cross-sections through horizontal plane(XY), sagittal plane (YZ), and coronal plane (XZ).A–C, 30 minutes at 4°C (inhibit internalization);D–F,30 minutes at 37°C; and G–I, 60 minutes at 37°C.

Doñate, F.,Raitano, A., Morrison, K., An, Z., Capo, L., Aviña, H., & Moriya, R.(2016).AGS16F is a novel antibody drug conjugate directed against ENPP3 for the treatment of renal cell carcinoma.Clinical Cancer Research,22(8),1989-1999.

Figure 3 AGS16F showed potent antitumor activity in RCC xenograft models.

Data are shown as mean ± SEM tumor volume for each group over time. A, mice (N = 10) bearing subcutaneous UG-K3 tumor xenografts (size ∼ 200 mm3) were treated with AGS16F by i.v. injection at doses of 0.125,0.25,and 0.5 mg/kg, every 4 days for 4 doses;B, mice bearing subcutaneous SKRC-01 cells tumor xenografts (size ∼ 200 mm3) were treated with single 5 or 10 mg/kg doses of AGS16F by i.v. injection; C, mice bearing subcutaneous RXF-393 cell tumor xenografts (size ∼ 100 mm3) were treated with single 3 or 10 mg/kg doses of AGS16F by i.v. injection.

Doñate, F.,Raitano, A., Morrison, K., An, Z., Capo, L., Aviña, H., & Moriya, R.(2016).AGS16F is a novel antibody drug conjugate directed against ENPP3 for the treatment of renal cell carcinoma.Clinical Cancer Research,22(8),1989-1999.

Figure 5 Mechanism of action of AGS16F and PD markers.

Mice bearing subcutaneous UG-K3 tumors (∼350 mm3) were treated as indicated, and biomarkers, ADC localization, and tumor volume were measured at 6, 24, 48, 72 hours, and 7 days. A, level of ccCK18 in serum over time. B, mean (±SEM) tumor volume for each group over time. C, percentage of FFPE tumor specimens positive for cleaved PARP-1 over time. D, percentage of FFPE tumor specimens positive for PPH3 over time. E,representative images of H&E staining of FFPE tumors at 24 hours and 7 days after dosing, with arrows showing cellular atypia and polymorphism. F, representative images of ENPP3 expression at 24 hours and 7 days after dosing.G, representative images of localization of AGS16F and control ADC in FFPE tumor samples using an antibody against MMAF at 24 hours and 7 days after dosing.

Doñate, F.,Raitano, A., Morrison, K., An, Z., Capo, L., Aviña, H., & Moriya, R.(2016).AGS16F is a novel antibody drug conjugate directed against ENPP3 for the treatment of renal cell carcinoma.Clinical Cancer Research,22(8),1989-1999.