Imperial College London
Alternate

Professor Karen Polizzi

Faculty of Engineering, Department of Chemical Engineering

Professor of Biotechnology
 
 
 
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Contact

 

+44 (0)20 7594 2851k.polizzi

 
 
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Location

 

411ACE ExtensionSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Spice:2020:10.1016/j.synbio.2020.06.003,
author = {Spice, AJ and Aw, R and Bracewell, DG and Polizzi, KM},
doi = {10.1016/j.synbio.2020.06.003},
journal = {Synthetic and Systems Biotechnology},
pages = {137--144},
title = {Improving the reaction mix of a Pichia pastoris cell-free system using a design of experiments approach to minimise experimental effort},
url = {http://dx.doi.org/10.1016/j.synbio.2020.06.003},
volume = {5},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - A renaissance in cell-free protein synthesis (CFPS) is underway, enabled by the acceleration and adoption of synthetic biology methods. CFPS has emerged as a powerful platform technology for synthetic gene network design, biosensing and on-demand biomanufacturing. Whilst primarily of bacterial origin, cell-free extracts derived from a variety of host organisms have been explored, aiming to capitalise on cellular diversity and the advantageous properties associated with those organisms. However, cell-free extracts produced from eukaryotes are often overlooked due to their relatively low yields, despite the potential for improved protein folding and posttranslational modifications. Here we describe further development of a Pichia pastoris cell-free platform, a widely used expression host in both academia and the biopharmaceutical industry. Using a minimised Design of Experiments (DOE) approach, we were able to increase the productivity of the system by improving the composition of the complex reaction mixture. This was achieved in a minimal number of experimental runs, within the constraints of the design and without the need for liquid-handling robots. In doing so, we were able to estimate the main effects impacting productivity in the system and increased the protein synthesis of firefly luciferase and the biopharmaceutical HSA by 4.8-fold and 3.5-fold, respectively. This study highlights the P. pastoris-based cell-free system as a highly productive eukaryotic platform and displays the value of minimised DOE designs.
AU  - Spice,AJ
AU  - Aw,R
AU  - Bracewell,DG
AU  - Polizzi,KM
DO  - 10.1016/j.synbio.2020.06.003
EP  - 144
PY  - 2020///
SN  - 2405-805X
SP  - 137
TI  - Improving the reaction mix of a Pichia pastoris cell-free system using a design of experiments approach to minimise experimental effort
T2  - Synthetic and Systems Biotechnology
UR  - http://dx.doi.org/10.1016/j.synbio.2020.06.003
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000561455200003&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - https://www.sciencedirect.com/science/article/pii/S2405805X20300302?via%3Dihub
UR  - http://hdl.handle.net/10044/1/83216
VL  - 5
ER  -
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