A new generation of E. coli expression hosts and tools for recombinant protein production
Lead Research Organisation:
Newcastle University
Department Name: Sch of Engineering
Abstract
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Planned Impact
As described in proposal submitted to TSB
Organisations
People |
ORCID iD |
| Phillip Craig Wright (Principal Investigator) |
| Description | Produced on an industrial scale, biotherapeutic proteins including cytokines, growth factors, and hormones, are of high importance and provide crucial therapeutic treatments. To understand how to better optimise the E. coli for this purpose, we here have applied quantitative proteomics tools iTRAQ and LFQ approaches to examine the differently expressed proteomes of two different mutated E. coli strains with targeted proteins (CyDisco proteins and Human Growth Hormone) grown under the presence and absence of IPTG (Isopropyl ß-D-1-thiogalactopyranoside). Two quantitative proteomics approaches have been used to address the question: How could CyDisco proteins regulate E. coli proteomes and recombinant protein. We were able to measure over 1300 proteins and to map these onto metabolic and regulatory pathways. The observations in this investigation provide useful information for understating the effect of CyDisco in order to improve recombinant protein production via cellular engineering. Papers are currently being written up. |
| Exploitation Route | We were able to generate a number of targets for metabolic and cellular engineering. These should provide options to manipulate E.coli. The data are with the project team at Kent for some of this cellular engineering. We were also able to understand options for purification of the target proteins. Again there are leads being explored at Kent and also at Birmingham. The industrial partners are also part of our discussions. |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | Starting to in the biopharmaceuticals manufacturing process. Understanding of cellular limitations in making these products. |
| First Year Of Impact | 2016 |
| Sector | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Title | MS - proteomics; iTRAQ |
| Description | Mass spectrometry method and informatics analysis. Not yet published. We plan to. |
| Type Of Material | Improvements to research infrastructure |
| Year Produced | 2017 |
| Provided To Others? | No |
| Impact | None as yet as we are still writing it up. |
| Description | Project meetings |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Industry/Business |
| Results and Impact | Engaging with academic and industry representatives on the outcomes of proteome analysis of E.coli cells producing biopharma products. Working out cellular bottlenecks and plans to limit these issues. Likely employment on more industrially relevant examples. |
| Year(s) Of Engagement Activity | 2015,2016,2017 |