Optimisation of CHO for Biotherapeutic Manufacture
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
Biological drugs (e.g. monoclonal antibodies, MAbs) based on recombinant DNA technology have transformed the treatment of life-limiting diseases including cancer, haemophilia and rheumatoid arthritis. The recent explosive growth in the biologics sector looks set to continue, with growing applications in precision medicine and personalised healthcare, and there are many new complex biologics in the drug discovery pipeline (e.g. bispecific, trispecific, and conjugated MAbs). The intrinsic complexity of these life-saving drugs is too challenging for synthesis by simple chemistry and requires the utilisation of living cells. Forcing cells to produce proteins that they do not naturally express is complex, and often requires a long period of trial and error cell manipulation, making the bio-manufacturing process time-consuming and very expensive and directly impacting on the delivery of transformative medicines to patients. With the recent remarkable development of powerful tools for editing mammalian genomes, new methods and automation for the synthesis of large numbers of DNA constructs, and the context provided by systems biology, the time is now right for using Synthetic Biology to establish a new paradigm for cost-effective manufacture of biologic drugs. In turn this will have a major impact on medicine and the health related industries, and make the biopharmaceutical value chain more cost-efficient.
The scale of the economic opportunity associated with this project is enormous. The UK has one of the most dynamic and innovative healthcare industries in the world and has developed over 20% of the world's top 100 selling drugs. The medical technology sector in the UK consists of around 2,800 companies, employing 52,000 people and generating around £10.6bn of turnover annually. An increasing portion of all medicines, currently estimated at 20%, are biopharmaceuticals. The global biologics market was valued at an estimated $251.5 billion in 2018 and is predicted to reach $319 billion by 2021. The CHO cell is the most widely used industrial expression system, which generates ~70% of approved and marketed therapeutic recombinant proteins, including multiple monoclonal antibodies (mAbs), so any enhancement of production efficiency and quality has a huge economic impact.
The vision of this prosperity partnership is to utilise state of the art investigational tools and synthetic biology approaches to both elucidate the intricacies of the CHO cell manufacturing platform and engineer it to be more predictive, effective, cost-efficient, and competitive for the production of biotherapeutics in the UK.
The scale of the economic opportunity associated with this project is enormous. The UK has one of the most dynamic and innovative healthcare industries in the world and has developed over 20% of the world's top 100 selling drugs. The medical technology sector in the UK consists of around 2,800 companies, employing 52,000 people and generating around £10.6bn of turnover annually. An increasing portion of all medicines, currently estimated at 20%, are biopharmaceuticals. The global biologics market was valued at an estimated $251.5 billion in 2018 and is predicted to reach $319 billion by 2021. The CHO cell is the most widely used industrial expression system, which generates ~70% of approved and marketed therapeutic recombinant proteins, including multiple monoclonal antibodies (mAbs), so any enhancement of production efficiency and quality has a huge economic impact.
The vision of this prosperity partnership is to utilise state of the art investigational tools and synthetic biology approaches to both elucidate the intricacies of the CHO cell manufacturing platform and engineer it to be more predictive, effective, cost-efficient, and competitive for the production of biotherapeutics in the UK.
Publications
Christofi E
(2023)
Ion Mobility Mass Spectrometry (IM-MS) for Structural Biology: Insights Gained by Measuring Mass, Charge, and Collision Cross Section.
in Chemical reviews
Donaldson J
(2022)
Synthetic biology approaches for dynamic CHO cell engineering.
in Current opinion in biotechnology
Donaldson JS
(2021)
Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach.
in Frontiers in bioengineering and biotechnology
Mentlak DA
(2024)
Dissecting cell death pathways in fed-batch bioreactors.
in Biotechnology journal
Moses T
(2023)
Right in two: capabilities of ion mobility spectrometry for untargeted metabolomics.
in Frontiers in molecular biosciences
Pennington O
(2024)
A multiscale hybrid modelling methodology for cell cultures enabled by enzyme-constrained dynamic metabolic flux analysis under uncertainty.
in Metabolic engineering
Samad SS
(2023)
Functional selectivity of Receptor Tyrosine Kinases regulates distinct cellular outputs.
in Frontiers in cell and developmental biology
Sizer RE
(2022)
TFIIIC-based chromatin insulators through eukaryotic evolution.
in Gene
Sizer RE
(2023)
Use of ubiquitous chromatin opening elements (UCOE) as tools to maintain transgene expression in biotechnology.
in Computational and structural biotechnology journal
Torres M
(2025)
Deciphering molecular drivers of lactate metabolic shift in mammalian cell cultures.
in Metabolic engineering
| Description | We have collected genome and transcriptome data for FDB's Apolloâ„¢X CHO cells, providing a comprehensive understanding of their genetic makeup and gene expression patterns. A JBrowse-based genome browser was built (hosted at UoE) and shared within the consortium with secure access. Proteomics and tRNAomic data has been collected. Metabolomics data has been collected from three mAb producing lines and a non-producing line for comparison. We have successfully developed various vectors and synthetic genetic devices for enhanced therapeutic protein production. "Sense-and-response" modules for ER stress mitigation have been created. Expression vector gene arrangement and orientation has been optimised and novel transposon-based vectors developed. CRISPR-based synthetic transcription factors and other expression enhancing vector elements are under development. tRNA gene barriers have been shown to boost productivity and cell line stability, findings have been recently published [doi:10.1002/biot.202400196]. Key cellular states have been characterised by metabolic and gene expression profiling under varying process formats - batch, fed-batch and perfusion culture. Several hypotheses have been tested by environmental and/or genetic intervention leading to enhance cell growth and antibody productivity. We have established metabolite replacement protocols, feeding regimes and metabolic requirements that control biomass and antibody productivity in diverse culture formats. Additionally, we identified the key activated cell death pathways in bioreactors, these findings have been published earlier this year [10.1002/biot.202300257]. Genetic interventions have led to enhanced glucose metabolic pathway regulation, increased efficiency of energy generation and establishment of means to switch the balance between cell growth and antibody productivity in a regulated manner. Data from the multi omics work packages have been used to identify target genes and pathways for engineering biology interventions. The iCHO2441 genome-scale metabolic model (GEM) [doi: 10.1002/bit.28366] was tailored for FDB's industrially relevant production system using extensive multi-omics data sets. Flux sampling was employed to identify metabolic signatures linked with high productivity phenotypes. These models were used to simulate media and feed supplementation strategies, validating predictions against experimental data and informing further media/feed supplements. |
| Exploitation Route | This is a prosperity partnership with Fujifilm Diosynth Biotechnologies. Results from the work will be implemented by Fujifilm in their manufacturing processes. |
| Sectors | Healthcare Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
| Description | The optimised expression technologies developed during this project have greatly enhanced biomanufacturing efficiency and FDB is actively commercialising these innovations: 1. A newly developed upstream process improves mAb product quality by ~15% (charge heterogeneity reduction) 2. A newly developed feed reduces raw material costs for bioreactor production by 50% 3. New expression vector components improve cell line stability and increases cellular productivity by >50%. The innovations resulting from this prosperity partnership have already had enormous commercial benefit for FDBK. These and future innovations will enhance the yield, quality and cost-effectiveness of biopharmaceutical production, strengthening the competitive position of FDB and the UK in the biopharmaceutical market globally. |
| First Year Of Impact | 2024 |
| Sector | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |