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The demands of the usage of hazardous ingredients for sulfur curing system in latex industries decrease with an increase in health-conscious and environmental awareness. This work demonstrates the incorporation of cassava starch (CS) as biodegradable fillers with natural rubber latex (NRL) through a sulfur-free crosslinking technique using radiation pre-vulcanization natural rubber latex (RVNRL) in comparison to sulfur pre-vulcanized natural rubber latex (PvNRL). The 20% CS dispersion was prepared, and 5–25 phr of dispersed CS content were compounded with NRL and formed into films by the coagulant dipping method. Microstructures and crystallinity of the films were analyzed by scanning electron microscopy (SEM) and X-ray diffraction, and their mechanical properties of NRL/CS films were characterized by tensile and tear tests. The result revealed that the crystallinity of RVNRL films was lower than PvNRL films. The total bond of S−C from PvNRL contributes to high tensile strength compared to C−C intermolecular rubber bond from radiation vulcanization system. The trend of decrement of tensile properties from sulfur crosslinking was larger than radiation crosslinking, and both systems gave similar tensile behavior at 25 phr of CS content. This attributed to the better dispersion of CS in RVNRL as confirmed by SEM micrographs. It was found that the optimum tear strength of RVNRL/CS and PvNRL/CS films was obtained at 10 and 5 phr of filler content, respectively. The result presented in this study may facilitate a contribution to the current literature on the development of latex film by radiation pre-vulcanization for rubber industry in the future. [ABSTRACT FROM AUTHOR]