Towards microfluidic reactors for cell-free protein synthesis at the point-of-care

Timm, Andrea C.; Shankles, Peter G.; Foster, Carmen M.; Doktycz, Mitchel John; Retterer, Scott T.

doi:10.1002/smll.201502764

Title: Towards microfluidic reactors for cell-free protein synthesis at the point-of-care

Journal Article · Tue Dec 22 00:00:00 EST 2015 · Small

DOI:https://doi.org/10.1002/smll.201502764· OSTI ID:1238743

Timm, Andrea C. ^[1]; Shankles, Peter G. ^[2]; Foster, Carmen M. ^[1]; Doktycz, Mitchel John ^[2]; Retterer, Scott T. ^[2]

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)

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 nano-fluidic architectures, CFPS can be optimized for point-of care use. Here, we describe 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. 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, prolonging the CFPS reaction and increasing protein yield. Membrane permeability 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. As a result, we show 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.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: Work for Others (WFO)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1238743

Journal Information:: Small, Vol. 12, Issue 6; ISSN 1613-6810

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 46 works

Citation information provided by
Web of Science

References (37)

Advances in Escherichia coli production of therapeutic proteins Swartz, James R. Current Opinion in Biotechnology, Vol. 12, Issue 2 https://doi.org/10.1016/S0958-1669(00)00199-3	journal	April 2001
Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements Huang, Chung-Jr; Lin, Henry; Yang, Xiaoming Journal of Industrial Microbiology & Biotechnology, Vol. 39, Issue 3 https://doi.org/10.1007/s10295-011-1082-9	journal	January 2012
Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines Kanter, Gregory; Yang, Junhao; Voloshin, Alexei Blood, Vol. 109, Issue 8 https://doi.org/10.1182/blood-2006-07-030593	journal	December 2006
Advances in cell-free protein synthesis for the functional and structural analysis of membrane proteins Junge, Friederike; Haberstock, Stefan; Roos, Christian New Biotechnology, Vol. 28, Issue 3 https://doi.org/10.1016/j.nbt.2010.07.002	journal	April 2011
High efficiency cell-free synthesis of proteins: Refinement of the coupled transcription/translation system Kudlicki, Wieslaw; Kramer, Gisela; Hardesty, Boyd Analytical Biochemistry, Vol. 206, Issue 2 https://doi.org/10.1016/0003-2697(92)90383-I	journal	November 1992
Prolonging cell-free protein synthesis with a novel ATP regeneration system Kim, Dong-Myung; Swartz, James R. Biotechnology and Bioengineering, Vol. 66, Issue 3 https://doi.org/10.1002/(SICI)1097-0290(1999)66:3<180::AID-BIT6>3.0.CO;2-S	journal	January 1999
Prolonging Cell-Free Protein Synthesis by Selective Reagent Additions Kim, D. -M.; Swartz, J. R. Biotechnology Progress, Vol. 16, Issue 3 https://doi.org/10.1021/bp000031y	journal	June 2000
Regeneration of adenosine triphosphate from glycolytic intermediates for cell-free protein synthesis Kim, Dong-Myung; Swartz, James R. Biotechnology and Bioengineering, Vol. 74, Issue 4 https://doi.org/10.1002/bit.1121	journal	January 2001
Mimicking theEscherichia coli cytoplasmic environment activates long-lived and efficient cell-free protein synthesis Jewett, Michael C.; Swartz, James R. Biotechnology and Bioengineering, Vol. 86, Issue 1 https://doi.org/10.1002/bit.20026	journal	January 2004
Substrate replenishment extends protein synthesis with an in vitro translation system designed to mimic the cytoplasm Jewett, Michael C.; Swartz, James R. Biotechnology and Bioengineering, Vol. 87, Issue 4 https://doi.org/10.1002/bit.20139	journal	January 2004
Rapid Expression and Purification of 100 nmol Quantities of Active Protein Using Cell-Free Protein Synthesis Jewett, Michael C.; Swartz, James R. Biotechnology Progress, Vol. 20, Issue 1 https://doi.org/10.1021/bp0341693	journal	January 2004
Synthesis of 2.3 mg/ml of protein with an all Escherichia coli cell-free transcription–translation system Caschera, Filippo; Noireaux, Vincent Biochimie, Vol. 99 https://doi.org/10.1016/j.biochi.2013.11.025	journal	April 2014
Preparative scale cell-free expression systems: New tools for the large scale preparation of integral membrane proteins for functional and structural studies Schwarz, Daniel; Klammt, Christian; Koglin, Alexander Methods, Vol. 41, Issue 4, p. 355-369 https://doi.org/10.1016/j.ymeth.2006.07.001	journal	April 2007
Expression of Active Murine Granulocyte-Macrophage Colony-Stimulating Factor in an Escherichia coli Cell-Free System Yang, J.; Kanter, G.; Voloshin, A. Biotechnology Progress, Vol. 20, Issue 6 https://doi.org/10.1021/bp034350b	journal	December 2004
Development of cell-free protein synthesis platforms for disulfide bonded proteins Goerke, Aaron R.; Swartz, James R. Biotechnology and Bioengineering, Vol. 99, Issue 2 https://doi.org/10.1002/bit.21567	journal	January 2007
Cell-free protein synthesis: Applications come of age Carlson, Erik D.; Gan, Rui; Hodgman, C. Eric Biotechnology Advances, Vol. 30, Issue 5 https://doi.org/10.1016/j.biotechadv.2011.09.016	journal	September 2012
A continuous cell-free translation system capable of producing polypeptides in high yield Spirin, A.; Baranov, V.; Ryabova, L. Science, Vol. 242, Issue 4882 https://doi.org/10.1126/science.3055301	journal	November 1988
Microscale to manufacturing scale-up of cell-free cytokine production-a new approach for shortening protein production development timelines Zawada, James F.; Yin, Gang; Steiner, Alexander R. Biotechnology and Bioengineering, Vol. 108, Issue 7 https://doi.org/10.1002/bit.23103	journal	March 2011
Characterization of extended channel bioreactors for continuous-flow protein production Timm, Andrea C.; Shankles, Peter G.; Foster, Carmen M. Journal of Vacuum Science & Technology B, Nanotechnology and Microelectronics: Materials, Processing, Measurement, and Phenomena, Vol. 33, Issue 6 https://doi.org/10.1116/1.4932155	journal	October 2015
Continuous protein production in nanoporous, picolitre volume containers Siuti, Piro; Retterer, Scott T.; Doktycz, Mitchel J. Lab on a Chip, Vol. 11, Issue 20 https://doi.org/10.1039/c1lc20462a	journal	January 2011
Cell-Sized confinement in microspheres accelerates the reaction of gene expression Kato, Ayako; Yanagisawa, Miho; Sato, Yuko T. Scientific Reports, Vol. 2, Issue 1 https://doi.org/10.1038/srep00283	journal	February 2012
Modular microfluidics for point-of-care protein purifications Millet, L. J.; Lucheon, J. D.; Standaert, R. F. Lab on a Chip, Vol. 15, Issue 8 https://doi.org/10.1039/C5LC00094G	journal	January 2015
Cell-free protein expression in a microchannel array with passive pumping Khnouf, Ruba; Beebe, David J.; Fan, Z. Hugh Lab Chip, Vol. 9, Issue 1 https://doi.org/10.1039/B808034H	journal	January 2009
Cell-Free Protein Synthesis in Microfluidic Array Devices Mei, Q.; Fredrickson, C. K.; Simon, A. Biotechnology Progress, Vol. 23, Issue 6 https://doi.org/10.1021/bp070133p	journal	December 2007
Substrate replenishment and byproduct removal improve yeast cell-free protein synthesis Schoborg, Jennifer A.; Hodgman, C. Eric; Anderson, Mark J. Biotechnology Journal, Vol. 9, Issue 5 https://doi.org/10.1002/biot.201300383	journal	January 2014
Prediction of diffusion coefficients of plasmids Prazeres, D. M. F. Biotechnology and Bioengineering, Vol. 99, Issue 4 https://doi.org/10.1002/bit.21626	journal	January 2008
A Novel Correlation for Protein Diffusion Coefficients Based on Molecular Weight and Radius of Gyration He, L. -Z.; Niemeyer, B. Biotechnology Progress, Vol. 19, Issue 2 https://doi.org/10.1021/bp0256059	journal	April 2003
Conformational dynamics of bacteriophage T7 DNA polymerase and its processivity factor, Escherichia coli thioredoxin Akabayov, B.; Akabayov, S. R.; Lee, S. -J. Proceedings of the National Academy of Sciences, Vol. 107, Issue 34 https://doi.org/10.1073/pnas.1010141107	journal	August 2010
A Semicontinuous Prokaryotic Coupled Transcription/Translation System Using a Dialysis Membrane Kim, D. -M.; Choi, C. -Y. Biotechnology Progress, Vol. 12, Issue 5 https://doi.org/10.1021/bp960052l	journal	October 1996
Fabrication optimization of a miniaturized array device for cell-free protein synthesis Khnouf, Ruba; Chapman, Benjamin D.; Hugh Fan, Z. ELECTROPHORESIS, Vol. 32, Issue 22 https://doi.org/10.1002/elps.201100376	journal	October 2011
Fabrication of nanoporous membranes for tuning microbial interactions and biochemical reactions Shankles, Peter G.; Timm, Andrea C.; Doktycz, Mitchel J. Journal of Vacuum Science & Technology B, Nanotechnology and Microelectronics: Materials, Processing, Measurement, and Phenomena, Vol. 33, Issue 6 https://doi.org/10.1116/1.4932671	journal	November 2015
Fiji: an open-source platform for biological-image analysis Schindelin, Johannes; Arganda-Carreras, Ignacio; Frise, Erwin Nature Methods, Vol. 9, Issue 7 https://doi.org/10.1038/nmeth.2019	journal	June 2012
Dose Conversion from Recombinant Human Erythropoietin to Darbepoetin alfa: Recommendations From Clinical Studies Scott, Shane D. Pharmacotherapy, Vol. 22, Issue 9 Part 2 https://doi.org/10.1592/phco.22.14.160S.33398	journal	September 2002
Comparison of 5 vs 10 μg/kg per day of GM-CSF following dose-intensified chemotherapy with cisplatin, etoposide, and ifosfamide in patients with advanced testicular cancer Bokemeyer, C.; Schmoll, H. -J.; Metzner, B. Annals of Hematology, Vol. 67, Issue 2 https://doi.org/10.1007/BF01788130	journal	August 1993
The side-effect profile of GM-CSF Stern, Angelika C.; Jones, T. C. Infection, Vol. 20, Issue 2 https://doi.org/10.1007/BF01705031	journal	March 1992
A Mass Spectrometric Method for Quantitation of Intact Insulin in Blood Samples Darby, Shauna M.; Miller, Mark L.; Allen, Ralph O. Journal of Analytical Toxicology, Vol. 25, Issue 1 https://doi.org/10.1093/jat/25.1.8	journal	January 2001
Recombinant activated clotting factor VII (rFVIIa) in the treatment of surgical and spontaneous bleeding episodes in hemophilic patients Ng, Heng Joo; Lee, Lai Heng Vascular Health and Risk Management, Vol. 2, Issue 4 https://doi.org/10.2147/vhrm.2006.2.4.433	journal	December 2006

Cited By (18)

Cell-free production of a therapeutic protein: Expression, purification, and characterization of recombinant streptokinase using a CHO lysate Tran, Kevin; Gurramkonda, Chandrasekhar; Cooper, Merideth A. Biotechnology and Bioengineering, Vol. 115, Issue 1 https://doi.org/10.1002/bit.26439	journal	October 2017
Cell‐free protein synthesis: The transition from batch reactions to minimal cells and microfluidic devices Ayoubi‐Joshaghani, Mohammad H.; Dianat‐Moghadam, Hassan; Seidi, Khaled Biotechnology and Bioengineering, Vol. 117, Issue 4 https://doi.org/10.1002/bit.27248	journal	January 2020
Tuning of salt separation efficiency by flow rate control in microfluidic dynamic dialysis Kalikavunkal, Prameen C.; Green, Nicolas G.; de Planque, Maurits R. R. Microfluidics and Nanofluidics, Vol. 23, Issue 10 https://doi.org/10.1007/s10404-019-2277-z	journal	September 2019
Interplay between materials and microfluidics Hou, Xu; Zhang, Yu Shrike; Santiago, Grissel Trujillo-de Nature Reviews Materials, Vol. 2, Issue 5 https://doi.org/10.1038/natrevmats.2017.16	journal	April 2017
On-chip manufacturing of synthetic proteins for point-of-care therapeutics Murphy, Travis W.; Sheng, Jiayuan; Naler, Lynette B. Microsystems & Nanoengineering, Vol. 5, Issue 1 https://doi.org/10.1038/s41378-019-0051-8	journal	March 2019
Cell-free gene expression: an expanded repertoire of applications Silverman, Adam D.; Karim, Ashty S.; Jewett, Michael C. Nature Reviews Genetics, Vol. 21, Issue 3 https://doi.org/10.1038/s41576-019-0186-3	journal	November 2019
High-yield production of “difficult-to-express” proteins in a continuous exchange cell-free system based on CHO cell lysates Thoring, Lena; Dondapati, Srujan K.; Stech, Marlitt Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-12188-8	journal	September 2017
Partitioning of hydrogels in 3D-printed microchannels Kim, Yong Tae; Bohjanen, Sara; Bhattacharjee, Nirveek Lab on a Chip, Vol. 19, Issue 18 https://doi.org/10.1039/c9lc00535h	journal	January 2019
Flexible on-demand cell-free protein synthesis platform based on a tube-in-tube reactor Zhou, Caijin; Lin, Xiaomei; Lu, Yuan Reaction Chemistry & Engineering, Vol. 5, Issue 2 https://doi.org/10.1039/c9re00394k	journal	January 2020
Integration of cell-free protein synthesis and purification in one microfluidic chip for on-demand production of recombinant protein Xiao, Xiao; Zhou, Yuan; Sun, Yuqiong Biomicrofluidics, Vol. 12, Issue 5 https://doi.org/10.1063/1.5042307	journal	September 2018
Cell-Free Synthetic Biology: Engineering Beyond the Cell Perez, Jessica G.; Stark, Jessica C.; Jewett, Michael C. Cold Spring Harbor Perspectives in Biology, Vol. 8, Issue 12 https://doi.org/10.1101/cshperspect.a023853	journal	October 2016
Streptokinase: An Efficient Enzyme in Cardiac Medicine Zia, Muhammad A. Protein & Peptide Letters, Vol. 27, Issue 2 https://doi.org/10.2174/0929866526666191014150408	journal	January 2020
Application of Cell-Free Protein Synthesis for Faster Biocatalyst Development Rolf, Jascha; Rosenthal, Katrin; Lütz, Stephan Catalysts, Vol. 9, Issue 2 https://doi.org/10.3390/catal9020190	journal	February 2019
Microfluidic Devices for Drug Delivery Systems and Drug Screening Damiati, Samar; Kompella, Uday; Damiati, Safa Genes, Vol. 9, Issue 2 https://doi.org/10.3390/genes9020103	journal	February 2018
Cell-Free Approaches in Synthetic Biology Utilizing Microfluidics Damiati, Samar; Mhanna, Rami; Kodzius, Rimantas Genes, Vol. 9, Issue 3 https://doi.org/10.3390/genes9030144	journal	March 2018
A User’s Guide to Cell-Free Protein Synthesis Gregorio, Nicole E.; Levine, Max Z.; Oza, Javin P. Methods and Protocols, Vol. 2, Issue 1 https://doi.org/10.3390/mps2010024	journal	March 2019
Cell-Free Synthetic Biology Platform for Engineering Synthetic Biological Circuits and Systems Jeong, Dohyun; Klocke, Melissa; Agarwal, Siddharth Methods and Protocols, Vol. 2, Issue 2 https://doi.org/10.3390/mps2020039	journal	May 2019
Application of cell-free protein synthesis for faster biocatalyst development Rolf, Jascha; Rosenthal, Katrin; Lütz, Stephan Technische Universität Dortmund https://doi.org/10.17877/de290r-19948	text	January 2019

Similar Records

Characterization of extended channel bioreactors for continuous-flow protein production

Journal Article · Fri Oct 02 00:00:00 EDT 2015 · Journal of Vacuum Science and Technology B · OSTI ID:1238743

Timm, Andrea C.; Shankles, Peter G.; Foster, Carmen M.; +2 more

Fabrication of nanoporous membranes for tuning microbial interactions and biochemical reactions

Journal Article · Wed Oct 21 00:00:00 EDT 2015 · Journal of Vacuum Science and Technology B · OSTI ID:1238743

Shankles, Peter G.; Timm, Andrea C.; Doktycz, Mitchel J.; +1 more

Modular microfluidics for point-of-care protein purifications

Journal Article · Thu Jan 01 00:00:00 EST 2015 · Lab on a Chip · OSTI ID:1238743

Millet, L. J.; Lucheon, J. D.; Standaert, R. F.; +2 more

Related Subjects

60 APPLIED LIFE SCIENCES

Title: Towards microfluidic reactors for cell-free protein synthesis at the point-of-care

Citation Formats

References (37)

Cited By (18)

Similar Records

Related Subjects