Bioprocess monitoring is used to track the progress of a cell culture and ensure that the product quality is maintained. Current schemes for monitoring metabolism rely on offline measurements of samples of the extracellular medium. However, in the era of synthetic biology, it is now possible to design and implement biosensors that consist of biological macromolecules and are able to report on the intracellular environment of cells. The use of fluorescent reporter signals allows non-invasive, non-destructive and online monitoring of the culture, which reduces the delay between measurement and any necessary intervention. The present mini-review focuses on protein-based biosensors that utilize FRET as the signal transduction mechanism. The mechanism of FRET, which utilizes the ratio of emission intensity at two wavelengths, has an inherent advantage of being ratiometric, meaning that small differences in the experimental set-up or biosensor expression level can be normalized away. This allows for more reliable quantitative estimation of the concentration of the target molecule. Existing FRET biosensors that are of potential interest to bioprocess monitoring include those developed for primary metabolites, redox potential, pH and product formation. For target molecules where a biosensor has not yet been developed, some candidate binding domains can be identified from the existing biological databases. However, the remaining challenge is to make the process of developing a FRET biosensor faster and more efficient.
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October 2013
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Conference Article|
September 23 2013
Opportunities for bioprocess monitoring using FRET biosensors
Antony Constantinou;
Antony Constantinou
*Department of Life Sciences, Imperial College London, London SW7 2AZ, U.K.
†Centre for Synthetic Biology and Innovation, Imperial College London, London SW7 2AZ, U.K.
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Karen M. Polizzi
Karen M. Polizzi
1
*Department of Life Sciences, Imperial College London, London SW7 2AZ, U.K.
†Centre for Synthetic Biology and Innovation, Imperial College London, London SW7 2AZ, U.K.
1To whom correspondence should be addressed (email k.polizzi@imperial.ac.uk).
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Biochem Soc Trans (2013) 41 (5): 1146–1151.
Article history
Received:
June 05 2013
Citation
Antony Constantinou, Karen M. Polizzi; Opportunities for bioprocess monitoring using FRET biosensors. Biochem Soc Trans 1 October 2013; 41 (5): 1146–1151. doi: https://doi.org/10.1042/BST20130103
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