In the present study, polymeric nanoparticles were prepared via ionic gelation of tripolyphosphate (TPP) and chitosan. The morphologies and characteristics of chitosan nanoparticles were determined by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared (FT-IR) spectroscopy, respectively, and their mean particle sizes and zeta potentials were obtained using DLS. FT-IR was confirmed tripolyphosphoric groups of TPP-linked with amine groups of chitosan in the nanoparticles. SEM was showed a spherical, smooth, and almost homogenous structure for the nanoparticles. The influence of many factors on the encapsulation of compsobuthus scorpion venom on the prepared chitosan nanoparticles (CS-NPs) was also evaluated. The optimum encapsulation efficiency (99.98%) and loading capacity (80.44%) were obtained by chitosan concentration of 2 mg/ml, chitosan to TPP mass ratio of 2, and compsobuthus scorpion venom initial concentration of 500 mg/mL. In vitro release of nanoparticles showed an initial burst release of approximately 20% in the first 8 hours, followed by a slow and steady rate release for about 72 hours. It could be concluded that the compsobuthus scorpion venom-loaded CS-NPs can be considered as an antigen delivery candidate.