Key reaction components affect the kinetics and performance robustness of cell-free protein synthesis reactions
dc.contributor.author | Banks, AM | |
dc.contributor.author | Whitfield, CJ | |
dc.contributor.author | Brown, SR | |
dc.contributor.author | Fulton, DA | |
dc.contributor.author | Goodchild, SA | |
dc.contributor.author | Grant, C | |
dc.contributor.author | Love, J | |
dc.contributor.author | Lendrem, DW | |
dc.contributor.author | Fieldsend, JE | |
dc.contributor.author | Howard, TP | |
dc.date.accessioned | 2023-04-25T09:28:15Z | |
dc.date.issued | 2021-12-13 | |
dc.date.updated | 2023-04-25T07:07:47Z | |
dc.description.abstract | Cell-free protein synthesis (CFPS) reactions have grown in popularity with particular interest in applications such as gene construct prototyping, biosensor technologies and the production of proteins with novel chemistry. Work has frequently focussed on optimising CFPS protocols for improving protein yield, reducing cost, or developing streamlined production protocols. Here we describe a statistical Design of Experiments analysis of 20 components of a popular CFPS reaction buffer. We simultaneously identify factors and factor interactions that impact on protein yield, rate of reaction, lag time and reaction longevity. This systematic experimental approach enables the creation of a statistical model capturing multiple behaviours of CFPS reactions in response to components and their interactions. We show that a novel reaction buffer outperforms the reference reaction by 400% and importantly reduces failures in CFPS across batches of cell lysates, strains of E. coli, and in the synthesis of different proteins. Detailed and quantitative understanding of how reaction components affect kinetic responses and robustness is imperative for future deployment of cell-free technologies. | en_GB |
dc.description.sponsorship | Engineering and Physical Sciences Research Council (EPSRC) | en_GB |
dc.format.extent | 218-229 | |
dc.format.medium | Electronic-eCollection | |
dc.identifier.citation | Vol. 20, pp. 218-229 | en_GB |
dc.identifier.doi | https://doi.org/10.1016/j.csbj.2021.12.013 | |
dc.identifier.grantnumber | EP/N026683/1 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/133007 | |
dc.identifier | ORCID: 0000-0003-0340-7431 (Love, John) | |
dc.identifier | ScopusID: 7202207902 (Love, John) | |
dc.identifier | ORCID: 0000-0002-0683-2583 (Fieldsend, Jonathan E) | |
dc.language.iso | en | en_GB |
dc.publisher | Elsevier / Research Network of Computational and Structural Biotechnology | en_GB |
dc.relation.url | https://www.ncbi.nlm.nih.gov/pubmed/35024094 | en_GB |
dc.relation.url | https://doi.org/10.25405/data.ncl.17041931 | en_GB |
dc.rights | © 2021 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). | en_GB |
dc.subject | Automation | en_GB |
dc.subject | Cell-free protein synthesis (CFPS) | en_GB |
dc.subject | Design of Experiments (DoE) | en_GB |
dc.subject | Robustness | en_GB |
dc.subject | Statistical engineering | en_GB |
dc.title | Key reaction components affect the kinetics and performance robustness of cell-free protein synthesis reactions | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2023-04-25T09:28:15Z | |
dc.identifier.issn | 2001-0370 | |
exeter.place-of-publication | Netherlands | |
dc.description | This is the final version. Available on open access from Elsevier via the DOI in this record | en_GB |
dc.description | Data statement: All data are available both in Source Data files associated with this publication, and at https://doi.org/10.25405/data.ncl.17041931 | en_GB |
dc.identifier.eissn | 2001-0370 | |
dc.identifier.journal | Computational and Structural Biotechnology Journal | en_GB |
dc.relation.ispartof | Comput Struct Biotechnol J, 20 | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | en_GB |
dcterms.dateAccepted | 2021-12-08 | |
dc.rights.license | CC BY | |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2021-12-13 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2023-04-25T09:25:50Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2023-04-25T09:28:19Z | |
refterms.panel | A | en_GB |
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Except where otherwise noted, this item's licence is described as © 2021 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).