Nanoscale drug carriers play an important role in regulating the delivery, permeability, and retention of the drugs. Although various carriers have been used to encapsulate anticancer drugs, natural biomaterials are of great benefit for delivery and controlled release of drugs. We used the electrospray deposition system to synthesize gliadin and gliadin-gelatin composite nanoparticles for delivery and controlled release of an anticancer drug (e.g., cyclophosphamide). The size profile and synthesis of nanoparticles was characterized by dynamic light scattering and X-ray diffractometry. Cyclophosphamide was gradually released from the gliadin nanoparticles for 48 h. In contrast, the gliadin-gelatin composite nanoparticles released cyclophosphamide in a rapid manner. Furthermore, we demonstrated that breast cancer cells cultured with cyclophosphamide-loaded 7% gliadin nanoparticles for 24 h became apoptotic, confirmed by Western blotting analysis. Therefore, the gliadin-based nanoparticle could be a powerful tool for delivery and controlled release of anticancer drugs.