Imperial College London

ProfessorPaulFreemont

Faculty of MedicineDepartment of Medicine

Chair in Protein Crystallography
 
 
 
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Contact

 

+44 (0)20 7594 5327p.freemont

 
 
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Location

 

259Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Goers:2017:10.1002/bit.26254,
author = {Goers, L and Ainsworth, C and Goey, CH and Kontoravdi and Freemont, PS and Polizzi, KM},
doi = {10.1002/bit.26254},
journal = {Biotechnology and Bioengineering},
pages = {1290--1300},
title = {Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production},
url = {http://dx.doi.org/10.1002/bit.26254},
volume = {114},
year = {2017}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Many high-value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimise the productivity of these cultures it is important to monitor cellular metabolism, for example the utilisation of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole-cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity and robustness. Importantly, our whole-cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole-cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalise on the wealth of genetic operons for metabolite sensing available in Nature for the development of other whole-cell biosensors.
AU - Goers,L
AU - Ainsworth,C
AU - Goey,CH
AU - Kontoravdi
AU - Freemont,PS
AU - Polizzi,KM
DO - 10.1002/bit.26254
EP - 1300
PY - 2017///
SN - 1097-0290
SP - 1290
TI - Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production
T2 - Biotechnology and Bioengineering
UR - http://dx.doi.org/10.1002/bit.26254
UR - http://hdl.handle.net/10044/1/44165
VL - 114
ER -