Recently, targeted medication delivery applications have effectively used self-assembled nanoparticles (NPs). In this study, poloxamer 407 and silk sericin protein were mixed in various proportions in the presence of dimethyl sulfoxide (DMSO) to create nine formulas of self-assembled nanostructures that could transport the hydrophobic anticancer medication paclitaxel (PTX). The produced NPs were then examined to determine their size distribution, percent of entrapment efficiency (EE%), morphology, compatibility and in vitro drug release studies. The selected formula was spherical and had a particle size (145 nm), a PDI of (0.25). and EE% of 82. The FT-IR data show that PTX and excipients are compatible, and the in vitro data show that PTX releases continuously over a 24-hour period. In order to successfully transport hydrophobic anticancer drugs (PTX) to target locations, this work proposes silk sericin protein as a substitute natural biomaterial for the creation of self-assembled NPs in conjunction with the presence of poloxamer 407.