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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{Goey:2018:10.1080/19420862.2018.1433977,
author = {Goey, CH and Bell, D and Kontoravdi, K},
doi = {10.1080/19420862.2018.1433977},
journal = {mAbs},
pages = {476--487},
title = {Mild hypothermic culture conditions impact residual host cell protein composition post-protein a chromatography},
url = {http://dx.doi.org/10.1080/19420862.2018.1433977},
volume = {10},
year = {2018}
}

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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Host cell proteins (HCPs) are endogenous impurities, and their proteolytic and binding properties can compromise the integrity, and, hence, the stability and efficacy of recombinant therapeutic proteins such as monoclonal antibodies (mAbs). Nonetheless, purification of mAbs currently presents a challenge because they often co-elute with certain HCP species during the capture step of protein A affinity chromatography. A Quality-by-Design (QbD) strategy to overcome this challenge involves identifying residual HCPs and tracing their source to the harvested cell culture fluid (HCCF) and the corresponding cell culture operating parameters. Then, problematic HCPs in HCCF may be reduced by cell engineering or culture process optimization. Here, we present experimental results linking cell culture temperature and post-protein A residual HCP profile. We had previously reported that Chinese hamster ovary cell cultures conducted at standard physiological temperature and with a shift to mild hypothermia on day 5 produced HCCF of comparable product titer and HCP concentration, but with considerably different HCP composition. In this study, we show that differences in HCP variety at harvest cascaded to downstream purification where different residual HCPs were present in the two sets of samples post-protein A purification. To detect low-abundant residual HCPs, we designed a looping liquid chromatography-mass spectrometry experiment with continuous expansion of a preferred, exclude, and targeted peptide list. Mild hypothermic cultures produced 20% more residual HCP species, especially cell membrane proteins, distinct from the control. Critically, we identified that half of the potentially immunogenic residual HCP species were different between the two sets of samples.
AU  - Goey,CH
AU  - Bell,D
AU  - Kontoravdi,K
DO  - 10.1080/19420862.2018.1433977
EP  - 487
PY  - 2018///
SN  - 1942-0862
SP  - 476
TI  - Mild hypothermic culture conditions impact residual host cell protein composition post-protein a chromatography
T2  - mAbs
UR  - http://dx.doi.org/10.1080/19420862.2018.1433977
UR  - http://hdl.handle.net/10044/1/63191
VL  - 10
ER  -