In this section
The Network aims to promote multi-disciplinary approaches to address challenging vaccine-related questions. This page contains a curated list of publications that highlight high-impact and collaborative approaches.
@article{Aw:2020:10.1002/cpps.115,
author = {Aw, R and Spice, AJ and Polizzi, K},
doi = {10.1002/cpps.115},
journal = {Current protocols in protein science},
title = {Methods for expression of recombinant proteins using a Pichia pastoris cell-free system},
url = {http://dx.doi.org/10.1002/cpps.115},
volume = {102},
year = {2020}
}
TY - JOUR
AB - Cellfree protein synthesis is a powerful tool for engineering biology and has been utilized in many diverse applications, from biosensing and protein prototyping to biomanufacturing and the design of metabolic pathways. By exploiting host cellular machinery decoupled from cellular growth, proteins can be produced in vitro both on demand and rapidly. Eukaryotic cellfree platforms are often neglected due to perceived complexity and low yields relative to their prokaryotic counterparts, despite providing a number of advantageous properties. The yeast Pichia pastoris (also known as Komagataella phaffii) is a particularly attractive eukaryotic host from which to generate cellfree extracts, due to its ability to grow to high cell densities with high volumetric productivity, genetic tractability for strain engineering, and ability to perform posttranslational modifications. Here, we describe methods for conducting cellfree protein synthesis using P. pastoris as the host, from preparing the cell lysates to protocols for both coupled and linked transcriptiontranslation reactions. By providing these methodologies, we hope to encourage the adoption of the platform by new and experienced users alike.
AU - Aw,R
AU - Spice,AJ
AU - Polizzi,K
DO - 10.1002/cpps.115
PY - 2020///
SN - 1934-3655
TI - Methods for expression of recombinant proteins using a Pichia pastoris cell-free system
T2 - Current protocols in protein science
UR - http://dx.doi.org/10.1002/cpps.115
UR - http://hdl.handle.net/10044/1/83872
VL - 102
ER -
Clinical Senior Lecturer
Dr Christopher Chiu
vaccine.network@imperial.ac.uk
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