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Research

Biological systems - including the simplest cells - exhibit a broad range of functions to thrive in their environment. Research in the Imperial College Centre for Synthetic Biology is focused on the possibility of engineering the underlying biochemical processes to solve many of the challenges facing society, from healthcare to sustainable energy. In particular, we model, analyse, design and build biological and biochemical systems in living cells and/or in cell extracts, both exploring and enhancing the engineering potential of biology.

As part of our research we develop novel methods to accelerate the celebrated Design-Build-Test-Learn synthetic biology cycle. As such research in the Centre for Synthetic Biology highly multi- and interdisciplinary covering computational modelling and machine learning approaches; automated platform development and genetic circuit engineering ; multi-cellular and multi-organismal interactions, including gene drive and genome engineering; metabolic engineering; in vitro/cell-free synthetic biology; engineered phages and directed evolution; and biomimetics, biomaterials and biological engineering.

Publications

Citation

BibTex format

@article{Aw:2018:10.1016/j.pep.2018.03.013,
author = {Aw, R and McKay, P and Shattock, R and Polizzi, K},
doi = {10.1016/j.pep.2018.03.013},
journal = {Protein Expression and Purification},
pages = {43--50},
title = {A systematic analysis of the expression of the anti-HIV VRC01 antibody  in Pichia pastoris through signal peptide optimization},
url = {http://dx.doi.org/10.1016/j.pep.2018.03.013},
volume = {149},
year = {2018}
}

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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Pichia pastoris (Komagataella phaffi) has been used for recombinant protein production for over 30 years with over 5000 proteins reported to date. However, yields of antibody are generally low. We have evaluated the effect of secretion signal peptides on the production of a broadly neutralizing antibody (VRC01) to increase yield. Eleven different signal peptides, including the murine IgG1 signal peptide, were combinatorially evaluated for their effect on antibody titer. Strains using different combinations of signal peptides were identified that secreted approximately 2-7 fold higher levels of VRC01 than the previous best secretor, with the highest yield of 6.50 mg L-1 in shake flask expression. Interestingly it was determined that the highest yields were achieved when the murine IgG1 signal peptide was fused to the light chain, with several different signal peptides leading to high yield when fused to the heavy chain. Finally, we have evaluated the effect of using a 2A signal peptide to create a bicistronic vector in the attempt to reduce burden and increase transformation efficiency, but found it to give reduced yields compared to using two independent vectors.
AU  - Aw,R
AU  - McKay,P
AU  - Shattock,R
AU  - Polizzi,K
DO  - 10.1016/j.pep.2018.03.013
EP  - 50
PY  - 2018///
SN  - 1046-5928
SP  - 43
TI  - A systematic analysis of the expression of the anti-HIV VRC01 antibody  in Pichia pastoris through signal peptide optimization
T2  - Protein Expression and Purification
UR  - http://dx.doi.org/10.1016/j.pep.2018.03.013
UR  - http://hdl.handle.net/10044/1/58612
VL  - 149
ER  -