A novel horseradish peroxidase-encapsulated silica nanoparticle (SNP) was generated in this study under relatively mild conditions. The generated enzyme-encapsulated SNP were relatively uniform in size (average 70 ± 14.3 nm), monodispersed, and spherical, as characterized by transmission electron microscopy and scanning electron microscopy. The horseradish peroxidase encapsulated in silica nanoparticle exhibits biological properties, such as a pHdependent activity profile and km value, similar to that of free enzymes. Furthermore, enzyme-encapsulated SNP exhibited good operational stability for the repetitive usage with a relative standard deviation of 5.1 % (n = 10) and a high stability for long term storage (>60 days) at 4 °C. The feasibility of using enzyme-encapsulated SNP in prodrug cancer therapy was also demonstrated by its capability to convert the prodrug indole-3-acetic acid into cytotoxic peroxyl radicals and trigger the death of tumor cells. These results indicate that the developed enzyme-encapsulated SNP has potential in the applications of prodrug cancer therapy.