Abstract Here, we present a novel method of growing protein crystals with nanofibers. Protein crystals were grown by incorporating nanofibers. No obvious differences were observed in diffraction data between fiber-containing and as-grown crystals. The fiber-containing crystals displayed an increased tolerance to osmotic shock caused by soaking in 25% ethanol or 40% dimethyl sulfoxide. This means that the method allowed us to easily increase the crystal mechanical stability. Because the method is very simple, it will provide a variety of possibilities for protein crystallization.