Greener, more sustainable platforms for high-value recombinant protein production
Lead Research Organisation:
University of Kent
Department Name: Sch of Biosciences
Abstract
The greenerRPP project aims to markedly enhance the EU's capacity for the production of secreted high-quality recombinant
proteins, especially industrial enzymes and therapeutic proteins (biotherapeutics). The combined markets are in excess of $330 billion
p.a., and the products are of massive significance for healthcare and a sustainable bio-based economy. greenerRPP will address
urgent problems in these industries, such as difficult-to-express proteins and the rapidly-emerging issue of 'sustainability' in the
European biotechnology industry. It will deliver improvements in yield and process design that increase the industry's effectiveness
while simultaneously reducing high current levels of waste. It will deliver industrial enzymes that reduce energy and raw material
costs in countless industrial and domestic settings. To achieve this, we will develop powerful new microbial and mammalian cell
platforms that produce recombinant proteins in unusually high quantities and in unparalleled quality, and bring to market a suite of
super-producing platforms.
A comprehensive training programme will equip greenerRPP ESRs with the interdisciplinary and intersectoral skills required to be
future leaders in these industries, while emphasising sustainability and gender-diversity dimension principles at every level. Finally,
the programme addresses the stark global inequality in this biotechnology sector: recombinant protein production is largely carried
out in (or outsourced by) high-income countries, placing low/middle income countries at a massive disadvantage. greenerRPP
includes a series of South East Asian Institutions as Associated Partners, to develop a global network with a knowledge exchangesecondment
programme that boosts these countries' own manufacturing programmes. greenerRPP ESRs will thus enhance a
European industrial sector that has never been more important to society, while making it 'greener' and reaching out to lower income
countries.
proteins, especially industrial enzymes and therapeutic proteins (biotherapeutics). The combined markets are in excess of $330 billion
p.a., and the products are of massive significance for healthcare and a sustainable bio-based economy. greenerRPP will address
urgent problems in these industries, such as difficult-to-express proteins and the rapidly-emerging issue of 'sustainability' in the
European biotechnology industry. It will deliver improvements in yield and process design that increase the industry's effectiveness
while simultaneously reducing high current levels of waste. It will deliver industrial enzymes that reduce energy and raw material
costs in countless industrial and domestic settings. To achieve this, we will develop powerful new microbial and mammalian cell
platforms that produce recombinant proteins in unusually high quantities and in unparalleled quality, and bring to market a suite of
super-producing platforms.
A comprehensive training programme will equip greenerRPP ESRs with the interdisciplinary and intersectoral skills required to be
future leaders in these industries, while emphasising sustainability and gender-diversity dimension principles at every level. Finally,
the programme addresses the stark global inequality in this biotechnology sector: recombinant protein production is largely carried
out in (or outsourced by) high-income countries, placing low/middle income countries at a massive disadvantage. greenerRPP
includes a series of South East Asian Institutions as Associated Partners, to develop a global network with a knowledge exchangesecondment
programme that boosts these countries' own manufacturing programmes. greenerRPP ESRs will thus enhance a
European industrial sector that has never been more important to society, while making it 'greener' and reaching out to lower income
countries.
People |
ORCID iD |
| Colin Robinson (Principal Investigator) |
| Description | Prototype cells/mammalian cell strains with minimized stress responses We have taken three approaches to generate new engineered Chinese hamster ovary (CHO) cell factories to minimise stress responses. These are; (1) engineering of the CHO cell chassis to reduce stress on the secretory pathway, (2) engineering of the CHO cell to limit competition during recombinant biotherapeutic protein drug production, and (3) enhanced culturing bioprocesses based around feeding of important precursors in lipid metabolism to reduce stress placed upon energy and secretory demands of lipid metabolism during recombinant protein production. In summary, all of these approaches have resulted in increased recombinant protein yields. Engineering of the secretory capacity of the CHO-S cell We have engineered the CHO-S host cell to over-express a target gene that enhances the secretory capacity of the cells and reduces the stress placed upon the cell by a recombinant protein load. We have shown that the stress reduced, enhanced secretory capacity engineered CHO-S cells enhances the secretory expression of a model reporter protein approximately 4-fold and, in preliminary studies, of the recombinant biotherapeutic protein EPO by much more than this. Engineering lipid metabolism of CHOK1 host cells alongside enhanced culture feeding bioprocesses to reduce stress and enhance recombinant protein production We have also looked to reduce the secretory stress on the CHOK1 chassis by engineering lipid metabolism and developing associated culture feeding strategies. We have so far managed to change the lipid metabolism of such cells in a new approach and are now evaluating the impact of this on the cells ability to make biotherapeutic protein drugs. |
| Exploitation Route | We envisage that these findings will be of interest to those companies wishing to produce biotherapeutic protein drugs at higher yield and quality with reduced environmental impact. This will allow for the demands for such drugs to be met, potentially reducing the cost to the end user. |
| Sectors | Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
| Description | We have used the findings for this award in engagement activities to demonstrate the importance of technologies for the production of life-saving protein based medicines. This has helped inform others about the importance of such technologies and the need for engineering biology approaches to develop more sustainable approaches for their production. |
| First Year Of Impact | 2024 |
| Sector | Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Societal |
| Description | CHO cell expression and profiling |
| Organisation | University of Greifswald |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | We have helped train PhD students in how the undertake molecular biology cloning of sequences into mammalian cell expression vectors and then use these to generate CHO cell lines expressing the biotherapeutic proteins of interest, specifically monoclonal antibodies. We hosted two students in the laboratory for 3-4 months who have since returned to their own laboratories to transfer the knowledge. We maintain a working relationship with the groups involved. |
| Collaborator Contribution | The project partners have undertaken proteomic analysis on CHO cell samples of mammalian cells we have subjected to engineering biology approaches to improve their ability to produce biotherapeuitc protein drugs. The data has been returned to us and we continue to work with them to identify key targets for further manipulation. |
| Impact | Multi-disciplinary involving engineers, biologists and technologists. |
| Start Year | 2023 |
| Description | CHO cell expression and profiling |
| Organisation | University of Groningen |
| Country | Netherlands |
| Sector | Academic/University |
| PI Contribution | We have helped train PhD students in how the undertake molecular biology cloning of sequences into mammalian cell expression vectors and then use these to generate CHO cell lines expressing the biotherapeutic proteins of interest, specifically monoclonal antibodies. We hosted two students in the laboratory for 3-4 months who have since returned to their own laboratories to transfer the knowledge. We maintain a working relationship with the groups involved. |
| Collaborator Contribution | The project partners have undertaken proteomic analysis on CHO cell samples of mammalian cells we have subjected to engineering biology approaches to improve their ability to produce biotherapeuitc protein drugs. The data has been returned to us and we continue to work with them to identify key targets for further manipulation. |
| Impact | Multi-disciplinary involving engineers, biologists and technologists. |
| Start Year | 2023 |
| Description | NIBRT collaboration |
| Organisation | National Institute of Bioprocessing Research and Training |
| Country | Ireland |
| Sector | Academic/University |
| PI Contribution | We are working with NIBRT to undertaken tRNA sequencing using Nanopore technology to develop greener CHO cell expression systems. We have generated new CHO cell systems that have been engineered to enhance recombinant protein production. |
| Collaborator Contribution | Expertise from NIBRT have aided in the development of experimental approaches and interpretation of data. Access to equipment, technology and bioinformatic support has been made available. |
| Impact | New CHO cells with different translational capacity that can be tuned for specific target recombinant mRNA sequences. |
| Start Year | 2023 |
| Description | Open Day Demonstrations and Lab Tours |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Public/other audiences |
| Results and Impact | Discussion and demonstration of production of recombinant biotherapeutic proteins made in CHO cells and the science underpinning the production of these life saving medicines. Tours of the laboratories undertaking research into this area and discussions around sustainability of production of such molecules. |
| Year(s) Of Engagement Activity | 2024,2025 |
| Description | Open Days |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Other audiences |
| Results and Impact | Talks to potential undergraduate students and parents on the work within the laboratory around CHO cell bioprocessing and production of recombinant proteins and vaccines. Demonstratuions of work being undertaken and Q&A sessions. |
| Year(s) Of Engagement Activity | 2023,2024 |
| Description | Talk to school leavers in Dublin, Ireland. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Schools |
| Results and Impact | Talk detailing the importance of biotherapeutic based protein drugs in the clinic and the challenges in manufacturing these. |
| Year(s) Of Engagement Activity | 2024 |