Manufacturing of protein-based biomaterials is gaining momentum in biomedical applications. In this chapter, we describe the procedures to create a versatile platform for the one-pot fabrication of different types of protein-based biomaterials by coupling the in vitro protein synthesis with the protein immobilization on solid materials in one-pot. To this aim, a set of plasmids and a battery of solid materials must be developed to guarantee the selective immobilization of the nascent protein on the surfaces, giving rise to functional biomaterials. This methodology also allows functionalizing materials with two or more proteins to increase the biomaterial's functionalities. Herein, this technology only requires the genomic information encoding the target protein, the desired solid material, and the cell-free extract containing the protein synthesis machinery. The cooperative action of all these elements turns out this portable technology as an innovative strategy for prototyping the fabrication of biomaterials and shortening their processing time.
Keywords: Cell-free protein synthesis; Homopeptide tag; In situ protein immobilization; Protein-functionalized biomaterials; Synthetic biology.