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Title: Characterization of extended channel bioreactors for continuous-flow protein production

Journal Article · · Journal of Vacuum Science and Technology B
DOI:https://doi.org/10.1116/1.4932155· OSTI ID:1238735
 [1];  [2];  [1];  [1];  [2]
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)

In this paper, protein based therapeutics are an important class of drugs, used to treat a variety of medical conditions including cancer and autoimmune diseases. Requiring continuous cold storage, and having a limited shelf life, the ability to produce such therapeutics at the point-of-care would open up new opportunities in distributing medicines and treating patients in more remote locations. Here, the authors describe the first steps in the development of a microfluidic platform that can be used for point-of-care protein synthesis. While biologic medicines, including therapeutic proteins, are commonly produced using recombinant deoxyribonucleic acid (DNA) technology in large batch cell cultures, the system developed here utilizes cell-free protein synthesis (CFPS) technology. CFPS is a scalable technology that uses cell extracts containing the biological machinery required for transcription and translation and combines those extracts with DNA, encoding a specific gene, and the additional metabolites required to produce proteins in vitro. While CFPS reactions are typically performed in batch or fed-batch reactions, a well-engineered reaction scheme may improve both the rate of protein production and the economic efficiency of protein synthesis reactions, as well as enable a more streamlined method for subsequent purification of the protein product—all necessary requirements for point-of-care protein synthesis. In this work, the authors describe a new bioreactor design capable of continuous production of protein using cell-free protein synthesis. The bioreactors were designed with three inlets to separate reactive components prior to on-chip mixing, which lead into a long, narrow, serpentine channel. These multiscale, serpentine channel bioreactors were designed to take advantage of microscale diffusion distances across narrow channels in reactors containing enough volume to produce a therapeutic dose of protein, and open the possibility of performing these reactions continuously and in line with downstream purification modules. Here, the authors demonstrate the capability to produce protein over time with continuous-flow reactions and examine basic design features and operation specifications fundamental to continuous microfluidic protein synthesis.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
Grant/Contract Number:
AC05-00OR22725; ERKCZ01
OSTI ID:
1238735
Alternate ID(s):
OSTI ID: 1421221
Journal Information:
Journal of Vacuum Science and Technology B, Vol. 33, Issue 6; ISSN 2166-2746
Publisher:
American Vacuum Society/AIPCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 8 works
Citation information provided by
Web of Science

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Cited By (4)

Integration of cell-free protein synthesis and purification in one microfluidic chip for on-demand production of recombinant protein journal September 2018
New generation of bioreactors that advance extracellular matrix modelling and tissue engineering journal October 2018
Toward Microfluidic Reactors for Cell-Free Protein Synthesis at the Point-of-Care journal December 2015
On-chip manufacturing of synthetic proteins for point-of-care therapeutics journal March 2019