Transarterial chemoembolization (TACE) and transarterial radioembolization (TARE) are promising treatments for unresectable liver tumours. Some recent studies suggested that combining TACE and TARE in one treatment course might improve treatment efficacy through synergistic cytotoxicity effects. Nonetheless, current formulations do not facilitate a combination of chemo‐ and radio‐embolic agents in one delivery system. Therefore, this study aimed to synthesise a hybrid biodegradable microsphere loaded with both radioactive agent, samarium‐153 (153Sm) and chemotherapeutic drug, doxorubicin (Dox) for potential radio‐chemoembolization of advanced liver tumours. 152Sm and Dox‐loaded polyhydroxybutyrate‐co‐3‐hydroxyvalerate (PHBV) microspheres were prepared using water‐in‐oil‐in‐water solvent evaporation method. The microspheres were then sent for neutron activation in a neutron flux of 2 × 1012 n/cm2/s. The physicochemical properties, radioactivity, radionuclide purity, 153Sm retention efficiency, and Dox release profile of the Dox‐153Sm‐PHBV microspheres were analysed. In addition, in vitro cytotoxicity of the formulation was tested using MTT assay on HepG2 cell line at 24 and 72 h. The mean diameter of the Dox‐153Sm‐PHBV microspheres was 30.08 ± 2.79 μm. The specific radioactivity was 8.68 ± 0.17 GBq/g, or 177.69 Bq per microsphere. The 153Sm retention efficiency was more than 99%, tested in phosphate‐buffered saline (PBS) and human blood plasma over 26 days. The cumulative release of Dox from the microspheres after 41 days was 65.21 ± 1.96% and 29.96 ± 0.03% in PBS solution of pH 7.4 and pH 5.5, respectively. The Dox‐153Sm‐PHBV microspheres achieved a greater in vitro cytotoxicity effect on HepG2 cells (85.73 ± 3.63%) than 153Sm‐PHBV (70.03 ± 5.61%) and Dox‐PHBV (74.06 ± 0.78%) microspheres at 300 μg/mL at 72 h. In conclusion, a novel biodegradable microspheres formulation loaded with chemotherapeutic drug (Dox) and radioactive agent (153Sm) was successfully developed in this study. The formulation fulfilled all the desired physicochemical properties of a chemo‐radioembolic agent and achieved better in vitro cytotoxicity on HepG2 cells. Further investigations are needed to evaluate the biosafety, radiation dosimetry, and synergetic anticancer properties of the formulation.