Vitis vinifera extract assisted silver nanoparticles (AgNPs) were biosynthesized that was confirmed primarily by color change and a sharp plasmon absorption band was found at 449 nm. Biosynthesized AgNPs shape was spherical and the particle size of 17 nm in average was confirmed by transmission electron microscopy (TEM) images. Functional groups of AgNPs were identified by Fourier transform infrared spectroscopy (FTIR). Streptococcus aureus was the most sensitive bacteria towards the assisted V. vinifera AgNPs as their growth was 90% inhibited at 100 g/mL concentration. That was also confirmed by the zone of inhibition study. Up to 96 h, no biofilm was observed for K. pneumoniae at 40 g/mL of AgNPs. Although AgNPs showed a mild toxicity against brine shrimp nauplii, it showed a remarkable level of antiproliferative activity against Ehrlich ascites carcinoma (EAC) cells.