The synthesis of silk protein nanoparticles entails a complex multiphase process. Selecting the most suitable production method is crucial for applying the resulting nanoparticles in biomedical fields and beyond. To achieve the ideal fibroin form for nanoparticle formulation, regardless of the chosen synthesis method, the silk fiber under goes two essential processes: degumming and dissolution, followed by dialysis. When selecting the method for fibroin nanoparticle formulation, an essential factor to consider is the ability to manipulate specific nanoparticle properties, such assize, shape, and zeta potential. Furthermore, the therapeutic efficacy of these nanoparticles is associated with their encapsulation rate, drug release profile, and biocompatibility. Among the various methods available for silk fibroin nanoparticle formulation, desolvation is the most commonly used, as it offers significant advantages, including ease of execution, adjustable size, and low operational costs. This method has resulted inconsiderable advancements in the targeted delivery of compounds for treating various types of cancer, such as breast cancer, glioblastoma and melanoma.