title: Toward Microfluidic Reactors for Cell-Free Protein Synthesis at the Point-of-Care.
abstract: Cell-free protein synthesis (CFPS) is a powerful technology that allows for optimization of protein production without maintenance of a living system. Integrated within micro and nanofluidic architectures, CFPS can be optimized for point-of-care use. Here, the development of a microfluidic bioreactor designed to facilitate the production of a single-dose of a therapeutic protein, in a small footprint device at the point-of-care, is described. This new design builds on the use of a long, serpentine channel bioreactor and is enhanced by integrating a nanofabricated membrane to allow exchange of materials between parallel "reactor" and "feeder" channels. This engineered membrane facilitates the exchange of metabolites, energy, and inhibitory species, and can be altered by plasma-enhanced chemical vapor deposition and atomic layer deposition to tune the exchange rate of small molecules. This allows for extended reaction times and improved yields. Further, the reaction product and higher molecular weight components of the transcription/translation machinery in the reactor channel can be retained. It has been shown that the microscale bioreactor design produces higher protein yields than conventional tube-based batch formats, and that product yields can be dramatically improved by facilitating small molecule exchange within the dual-channel bioreactor.
authors.affiliation: Oak Ridge National Laboratory, 1 Bethel Valley Road, Oak Ridge, TN, 37831, USA.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Timm
authors.affiliation: Oak Ridge National Laboratory, 1 Bethel Valley Road, Oak Ridge, TN, 37831, USA.|The University of Tennessee, Knoxville, TN, 37996, USA.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Shankles
authors.affiliation: Oak Ridge National Laboratory, 1 Bethel Valley Road, Oak Ridge, TN, 37831, USA.
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Foster
authors.affiliation: Oak Ridge National Laboratory, 1 Bethel Valley Road, Oak Ridge, TN, 37831, USA.|The University of Tennessee, Knoxville, TN, 37996, USA.
authors.forename: Mitchel J
authors.initials: MJ
authors.lastname: Doktycz
authors.affiliation: Oak Ridge National Laboratory, 1 Bethel Valley Road, Oak Ridge, TN, 37831, USA.|The University of Tennessee, Knoxville, TN, 37996, USA.
authors.forename: Scott T
authors.initials: ST
authors.lastname: Retterer
mesh_terms: D019149:Bioreactors; D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D008567:Membranes, Artificial; D044085:Microfluidics / Q000295:instrumentation*; D058608:Nanopores / Q000648:ultrastructure; D010539:Permeability; D019095:Point-of-Care Systems; D016062:Porosity; D014176:Protein Biosynthesis; D012825:Silicon / Q000737:chemistry
chemical_list: D008567:Membranes, Artificial; D012825:Silicon
keywords: biologics; cell-free protein synthesis; electron-beam lithography; microfluidics; microreactors