Translation of Step-changing Bioprocesses and Expression System Technologies for Next Generation Protein Biologics Production in CHO Cells

Lead Research Organisation: University of Kent
Department Name: Sch of Biosciences

Abstract

Chinese hamster ovary (CHO) cells are the main production host for >US$145billion/yr of protein biologics used as medicines for a range of diseases. The CHO platform is mature when considering production of monoclonal antibodies, but new format non-native molecules such as fusion proteins, antibody fragments and other exotic molecules remain difficult to express (DTE) in this, or any other host. This project builds upon proof of concept work demonstrating that engineering the CHO chassis, together with growth media manipulation, increases both the yield and quality of a number of DTE proteins that are in development for application to unmet clinical needs and diseases with no current treatments. The project will advance the technology readiness level of our preliminary findings beyond proof-of-concept to deliver the commercialization of new CHO cell systems for DTE proteins and associated bioprocesses ready for industrial application to produce these important new medicines.

Technical Summary

This proposal's overall output is a novel integrated pipeline for difficult to express (DTE) biologics production. A 3-fold approach to realising the commercial potential of the technology will be adopted. Firstly, a lipid suplementation strategy will be developed to improve existing CHO processes. Secondly, a suite of expression vectors will be created comprising new marker genes that complement CHO auxotrophies and lipid biosynthesis genes. Thirdly, engineered CHO chassis will be generated that stably express the lipid metabolism genes and/or combinations of these. The final, and preferred, embodiment of the technology will be a combination of new cell chassis and improved processes. This approach will maximise the realisation of value by ensuring early implmentation and broad application (both existing and new cell lines) and also mitigates against the failure of any one approach. WPs 1-4 are based upon innovations that both teams together were involved in developing. Specific milestones: 1. Appointment of staff/training (Q1, y1). 2. 3/4 gene cassette vectors available (Q2, y1). 3. Vectors with novel metabolic selection markers (tyrosine/proline) available (Q4, y1). 4. New lipid supplementation processes for DTE proteins in existing technologies developed (Q4, y1). 5. Initial lipid engineered Xceed host cell line available and validated for enhanced DTE protein yield/quality (Q2, y2). 6. Cell line construction and process development validated for novel lipid engineered Xceed hosts (Q2, y3). 7. cGMP cell banks available (Q2, y3). 8. cGMP cell bank testing complete, novel cell lines and upload of process conditions suitable for cGMP use into Lonza's GMP document management system and release of a process uptake to Lonza's GS Licensees (Q4, y3).

Planned Impact

As described in proposal submitted to IUK
 
Description This award is only approximately 75% of the way towards completion but has already yielded significant findings. We have developed new selection systems for the generation of stable recombinant Chinese hamster ovary cell lines that have been filed to be protected. We have also developed new metabolic selection markers and engineered CHO cell metabolism to overcome industrial challenges associated with metabolism. These new technologies are now being evaluated in the commercial environment. We have also established new synthetic plasmid DNA based vector systems for the expression of multiple genes in mammalian cell systems.The team has also now generated a range of potential new engineered CHO host cell lines to expand the capacity of these cells to produce difficult to express proteins. The impact of these manipulations is again being validated in the commercial environment. Finally, we have established methods for manipulating specific processes in CHO cells that results in enhancing their ability to make recombinant protein based medicines at greater yield and quality.
Exploitation Route Industrially for the commercial manufacture of protein based medicines.
Sectors Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description Although this project is early in its progression, already we have generated findings that are being validated and applied in an industrial setting for the manufacture of recombinant biotherapeutic proteins. These will be further evaluated and developed as the project progresses. In particular, we have developed engineering of CHO cells that help overcome some bioprocess challenges and provide a unique solution to these problems. These are now being validated in the commercial environment. We have also developed engineered cell lines to enhance the ability of such cells to produce difficult to express recombinant proteins. This has relied upon new selection technology developed during the project. The project is on-going.
First Year Of Impact 2016
Sector Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Description KTP Award
Amount £204,432 (GBP)
Funding ID KTP010320 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 07/2016 
End 01/2019
 
Description UCL Biochemical Engineering CDT PhD Award
Amount £58,212 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description IBCatalyst LMM collaboration 
Organisation Lonza Group
Country Global 
Sector Private 
PI Contribution This project has been in partnership with Lonza Biologics, Slough, UK to develop and commercialise new technology for the production of difficult to express recombinant proteins from Chinese hamster ovary (CHO) cells. Both partners have contributed knowledge and knowhow and expertise to deliver the project and objectives. The project is still on-going as of March 2019.
Collaborator Contribution As above.
Impact Commercially confidential at this stage. Patents submitted, publications will arise once these are granted.
Start Year 2016
 
Title INHIBITION OF PROTEIN DEGRADATION FOR IMPROVED PRODUCTION 
Description Disclosed herein are methods and compositions useful for evaluating, selecting, identifying, or making a cell or cell line that has improved production capacity for generating higher yields of products and/or improved capacity to produce higher quality products. Products, as described herein, can include a polypeptide that is endogenously expressed by the cell, a recombinant polypeptide that is not endogenously expressed, or a non-naturally occurring recombinant polypeptide. The methods described herein include modulating, e.g., inhibiting, the protein degradation pathway by using a proteasome inhibitor, an ER-associated degradation (ERAD) inhibitor, or a ubiquitin pathway inhibitor. 
IP Reference WO2017118726 
Protection Patent application published
Year Protection Granted 2017
Licensed Commercial In Confidence
Impact Understanding of how protein turnover can be used to select for Chinese hamster ovary (CHO) cells with an increased capacity to generate secretory recombinant proteins.
 
Title Modulation of lipid metabolism for protein production 
Description The present disclosure features methods and compositions for modulating lipid metabolism to achieve improved production and quality of recombinant products, such as next generation biologics. Modulation of lipid metabolism as described herein includes, for example, introducing a lipid metabolism modulator described herein to a cell or a cell-free system. Also encompassed by the present disclosure are engineered cells with improved production capacity and improved product quality, methods for engineering such cells, and preparations and mixtures comprising the products from such cells. 
IP Reference WO2017191165 
Protection Patent application published
Year Protection Granted 2016
Licensed Commercial In Confidence
Impact This has contributed towards knowledge around lipid metabolism in recombinant CHO producing cell lines and the importance of this for underpinning recombinant cell growth and productivity.
 
Description 7 open days at University 
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 Describing the research we undertake in the laboratory, covering all aspects of the research and the impact this has/can have. Particular questions around genetic modification of cells to produce recombinant proteins in all sessions and discussions around both the ethical aspects of this and the potential applications of such technology.
Year(s) Of Engagement Activity 2015,2016
 
Description Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at international BBSRC NIBB secretion meeting in Glasgow, Scotland.
Year(s) Of Engagement Activity 2017
 
Description Open Day Activities 
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 Activity involved practical demonstration of cell culture and gene editing. Also involved demonstration of mass spectrometry to determine post-translational modifications of proteins. Major activity was describing how engineering of cells can be used to produce recombinant biotherapeutic drugs.
Year(s) Of Engagement Activity 2018
 
Description Open day tours of research laboratory and discussions with participants 
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 Undergraduate students
Results and Impact 8 days of open day activities.
Year(s) Of Engagement Activity 2017
 
Description Presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at 13th Protein Expression in Animal Cells (PDRA on project)
Year(s) Of Engagement Activity 2017
URL http://www.peace-conference.org
 
Description Royal Society Summer Science Exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact The Summer Science Exhibition is an annual display of the most cutting-edge science and technology in the UK. This free, week-long festival features exhibits and a series of inspiring talks and activities for all ages. Meet the scientists, discover the exciting research and technology they work on and have fun with great hands-on activities.
Year(s) Of Engagement Activity 2018
URL https://royalsociety.org/science-events-and-lectures/2019/summer-science-exhibition/