This protocol describes the fabrication of silk-based biomaterials from Bombyx mori silk fibroin. Silk fibroin, derived from silkworm cocoons, is a versatile protein polymer with excellent mechanical properties, biocompatibility, and controllable degradation rates. The process involves extracting silk fibroin from cocoons, removing sericin, and then fabricating various materials such as hydrogels, tubes, sponges, composites, fibers, microspheres, and thin films. The extracted silk fibroin is dissolved in lithium bromide, dialyzed, and centrifuged to obtain a pure solution. This solution can be used directly or concentrated for further processing. The protocol includes methods for creating different material formats, such as dip methods for thin-walled tubes, gel spinning for tubes with texture, and various techniques for hydrogelation, film formation, microsphere production, and electrospinning of fibers. The materials can be sterilized and stored for future use. The process also highlights the importance of controlling factors such as temperature, time, and solution concentration to achieve desired material properties. The protocol provides detailed steps for each material fabrication method, along with necessary reagents and equipment. The final materials can be used in biomedical applications such as implants, tissue engineering, drug delivery, and disease modeling.This protocol describes the fabrication of silk-based biomaterials from Bombyx mori silk fibroin. Silk fibroin, derived from silkworm cocoons, is a versatile protein polymer with excellent mechanical properties, biocompatibility, and controllable degradation rates. The process involves extracting silk fibroin from cocoons, removing sericin, and then fabricating various materials such as hydrogels, tubes, sponges, composites, fibers, microspheres, and thin films. The extracted silk fibroin is dissolved in lithium bromide, dialyzed, and centrifuged to obtain a pure solution. This solution can be used directly or concentrated for further processing. The protocol includes methods for creating different material formats, such as dip methods for thin-walled tubes, gel spinning for tubes with texture, and various techniques for hydrogelation, film formation, microsphere production, and electrospinning of fibers. The materials can be sterilized and stored for future use. The process also highlights the importance of controlling factors such as temperature, time, and solution concentration to achieve desired material properties. The protocol provides detailed steps for each material fabrication method, along with necessary reagents and equipment. The final materials can be used in biomedical applications such as implants, tissue engineering, drug delivery, and disease modeling.