Imperial College London
Alternate

Professor Oscar Ces

Faculty of Natural Sciences, Department of Chemistry

Head of Department, August von Hofmann Chair of Chemistry
 
 
 
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Contact

 

+44 (0)20 7594 3754o.ces Website

 
 
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Location

 

G04AMolecular Sciences Research HubWhite City Campus

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Summary

 

Publications

Citation

BibTex format

@article{Heide:2021:10.3389/fbioe.2020.604091,
author = {Heide, C and Buldum, G and Moya-Ramirez, I and Ces, O and Kontoravdi, K and Polizzi, K},
doi = {10.3389/fbioe.2020.604091},
journal = {Frontiers in Bioengineering and Biotechnology},
title = {Design, development and optimisation of a functional mammalian cell-free protein synthesis platform},
url = {http://dx.doi.org/10.3389/fbioe.2020.604091},
volume = {8},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - In this paper, we describe the stepwise development of a cell-free protein synthesis (CFPS) platform derived from cultured Chinese hamster ovary (CHO) cells. We provide a retrospective summary of the design challenges we faced, and the optimized methods developed for the cultivation of cells and the preparation of translationally active lysates. To overcome low yields, we developed procedures to supplement two accessory proteins, GADD34 and K3L, into the reaction to prevent deactivation of the translational machinery by phosphorylation. We compared different strategies for implementing these accessory proteins including two variants of the GADD34 protein to understand the potential trade-offs between yield and ease of implementation. Addition of the accessory proteins increased yield of turbo Green Fluorescent Protein (tGFP) by up to 100-fold depending on which workflow was used. Using our optimized protocols as a guideline, users can successfully develop their own functional CHO CFPS system, allowing for broader application of mammalian CFPS.
AU  - Heide,C
AU  - Buldum,G
AU  - Moya-Ramirez,I
AU  - Ces,O
AU  - Kontoravdi,K
AU  - Polizzi,K
DO  - 10.3389/fbioe.2020.604091
PY  - 2021///
SN  - 2296-4185
TI  - Design, development and optimisation of a functional mammalian cell-free protein synthesis platform
T2  - Frontiers in Bioengineering and Biotechnology
UR  - http://dx.doi.org/10.3389/fbioe.2020.604091
UR  - http://hdl.handle.net/10044/1/86443
VL  - 8
ER  -
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