title: Continuous protein production in nanoporous, picolitre volume containers.
abstract: The synthetic manufacture of functional proteins enables a bottom-up understanding of the workings of biological systems and opens new opportunities for the treatment of disease. Cell-free protein synthesis is a practical approach for enabling such manufacturing, however, it is typically carried out in fairly large volumes, when compared to a natural cell, leading to increases in cost and loss of efficiency. Here we demonstrate continuous cell free protein synthesis in arrays of cellular scale containers that continuously exchange energy and materials with their environment. A multiscale fabrication process allows the monolithic integration of nanoporous silicon containers within an addressable microfluidic network. Synthesis of enhanced green fluorescent protein (eGFP) in the containers continues beyond 24 h and yields more than twice the amount of protein, on a per volume basis, than conventional scale batch reactions. By mimicking the physical volume and controlled flux of a natural cell, the resulting "cell mimic" devices can enable fundamental studies of biological systems as well as serve applications related to the functional screening of proteins and the on-demand production of biologics.
authors: Genome, Science and Technology Program, University of Tennessee, Knoxville, TN 37996, USA. psiuti@utk.edu
authors: Piro
authors: P
authors: Siuti
authors: Scott T
authors: ST
authors: Retterer
authors: Mitchel J
authors: MJ
authors: Doktycz
mesh_terms: D032701:Biomimetics / Q000295:instrumentation*; D002474:Cell-Free System; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D046210:Microfluidic Analytical Techniques; D058608:Nanopores; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins