Treatment with hSD5-vedotin at a dose of 1 mg/kg resulted in a 56 % tumor growth inhibition (TGI): The freshly cultured BxPc-3 cancer cells were harvested during the logarithmic growth phase, suspended in PBS, and subcutaneously implanted into NOD/SCID mice (5 × 106 cells per mouse) to induce tumor formation. Tumor size was measured bi-weekly, and the volume was calculated using the formula V = 0.5lw2, where l represents the length and w represents the width of the tumor. When the tumor size reached approximately 100 mm3, the animals were divided into groups that received the indicated treatments (1, 0.2, 0.04 mg/kg, iv, qwk). The mice were monitored frequently for any signs of adverse drug-related side effects.

The proliferation of PDAC cells was measured using the 3- (4,5-dimethylthiazol-2-yl)-5- (3-carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium (MTS) cell proliferation assay kit (Promega). The cells were seeded in a 96-well culture plate for attachment. The hSD5-vedotin at various concentrations was added to the cell culture and incubated for 5 days. Finally, MTS and phenazine methosulfate solutions were added and set for development. After the SDS reagent was added to stop the reaction, the absorbance of each well was measured at OD 490 nm.