| EPSRC Reference: |
GR/R80827/01 |
| Title: |
The development of on-line flow cytometry for bio-process monitoring and control |
| Principal Investigator: |
Hewitt, Professor C |
| Other Investigators: |
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| Researcher Co-Investigators: |
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| Project Partners: |
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| Department: |
Chemical Engineering |
| Organisation: |
University of Birmingham |
| Scheme: |
Fast Stream |
| Starts: |
19 April 2002 |
Ends: |
18 August 2005 |
Value (£): |
58,430
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| EPSRC Research Topic Classifications: |
| Bioprocess Engineering |
Design of Process systems |
| Instrumentation Eng. & Dev. |
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| EPSRC Industrial Sector Classifications: |
| Pharmaceuticals and Biotechnology |
No relevance to Underpinning Sectors |
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| Panel History: |
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Summary on Grant Application Form |
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There is an immediate need within the pharmaceutical industry to develop on-line, analytical techniques to make measurements on single cells during a bio-process. The aim being to accurately quantify important parameters such as cell viability, protein synthesis or plasmid copy number, in real-time, that allow an informed approach to be taken to process control. Flow cytometry has the potential to provide this information now that microbial physiology and fluorescent stain behaviour have been understood. Previously, the deliberate measurement of the intrinsic light scattering of a microbial population has allowed the at-line quanitifcation of contaminating cells and of inclusion body (insoluable protein) formation. Further the use of mufti-parameter fluorescent staining techniques developed in our laboratory have lead to a functional classification of the physiological state of individual bacterial cells based on the presence or absence of an intact poles cytoplasmic membrane and the functionality of the metabolic pumps across it. It is therefore possible to resolve an individual cells physiological state beyond culturability enabling assessment of population heterogeneity. These techniques have been used for the atline study of a range of microbial fermentations. It has been shown that during the latter stages of high cell density fed-batch fermentations that there is a considerable fluctuation in cell physiological state, including viability. Indeed a drop in cell viability of about 20%, as characterised by cytoplasmic membrane depolarisation and permeability is not uncommon. These phenomena are important since fed-batch high cell density cultivation is the preferred industrial method for increasing the volumetric productivity of bacterial products eg. heterolegous recombinant proteins (AstraZeneca, Avecia) or plasmid for gene therapy (Cobra Therapeutics) and the presence of a large number of dead cells will have an obvious detrimental effect on this.Given this understanding it is proposed that this project will aim to develop on-line systems for improved bio-process monitoring and control.
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Key Findings |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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Potential use in non-academic contexts |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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Impacts |
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| Description |
This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk |
| Summary |
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| Date Materialised |
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Sectors submitted by the Researcher |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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| Project URL: |
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| Further Information: |
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| Organisation Website: |
http://www.bham.ac.uk |