A Combinatorial Approach to Enhance Production of Monoclonal Antibodies
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
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Technical Summary
CHO cells are the most widely used industrial system for producing recombinant therapeutic proteins, but can struggle to express and secrete large biologics such as monoclonal antibodies (mAbs) at sustained high levels. We will address this with a systematic programme of synthetic cell bioengineering that combines innovative approaches to concomitantly increase CHO cells' capacities to express and secrete mAbs. Our team combines complementary skill sets to enhance successive steps that are potentially rate-limiting for mAb production, including transgene expression, mRNA translation, intracellular trafficking, post-translational modification and secretion. We will combine a series of novel improvements in a holistic manner to generate CHO cell lines optimised to produce mAbs of great economic and therapeutic value. In so doing, the project will establish a robust, flexible and adaptable UK platform for optimisation and manufacture of therapeutic biologics.
Planned Impact
As described in proposal submitted to TSB
People |
ORCID iD |
| Susan Rosser (Principal Investigator) |
Publications
Bryson JW
(2022)
Multiplexed activation in mammalian cells using a split-intein CRISPR/Cas12a based synthetic transcription factor.
in Nucleic acids research
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
Kleinjan DA
(2017)
Drug-tunable multidimensional synthetic gene control using inducible degron-tagged dCas9 effectors.
in Nature communications
Merrick CA
(2018)
Serine Integrases: Advancing Synthetic Biology.
in ACS synthetic biology
Olorunniji FJ
(2019)
Control of ?C31 integrase-mediated site-specific recombination by protein trans-splicing.
in Nucleic acids research
| Description | We have demonstrated - Addition of insulator cassette based on tRNA gene sequences positively impacts on productivity. Synthetic transcription factors based on a dCas9 scaffold have been shown to be effective at regulating gene expression in CHO cells. The LEAP system has been shown to be highly effective at rapid positive clone selection. Loci in the CHO genome have been identified for stability and high expression of transgenes Landing pads have been developed and gene insertion, deletion and replacement have been demonstrated. |
| Exploitation Route | Fujifilm Diosynth Biotechnologies UK (FDBK) have funded a Centre of Excellence (£2M) in protein production with the Universities of Edinburgh, York and Manchester with Prof Rosser as academic lead. This is leveraged on the work of this project. FDBK has been trialling the tRNA based insulator sequences in their commercial production cell lines UCB have funded a Case studentship to work on this project. We have recently been funded by the EPSRC for a prosperity partnership to with Fujifilm Diosynth Biotechnologies UK Patents have been filed on the insulators and the CHO genomic insertion sites. |
| Sectors | Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | Fujifilm Diosynth Biotechnologies UK (FDBK) have funded a Centre of Excellence (£2M) in protein production with the Universities of Edinburgh, York and Manchester with Prof Rosser as academic lead. This is leveraged on the work of this project. We have also recently had an EPSRC prosperity partnership funded with FDBK to further translate this work. FDBK has been trialling the tRNA based insulator sequences in their commercial production cell lines A patent has been filed on high producing CHO genomic insertion sites which has generated interest from a number of companies. In 2021 the EPSRC funded a large £7.8M Prosperity Partnership led by FDBK and the University of Edinburgh on the optimisation of CHO cells for biotherapeutic manufacture |
| First Year Of Impact | 2020 |
| Sector | Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Description | BBSRC IAA University of York |
| Amount | £300,000 (GBP) |
| Funding ID | BB/S506795/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 03/2018 |
| End | 03/2022 |
| Description | Development of a computational glycan engineering tool for biologics manufacturers |
| Amount | £198,765 (GBP) |
| Funding ID | BB/T016965/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 01/2021 |
| End | 12/2023 |
| Description | Optimisation of CHO for Biotherapeutic Manufacture |
| Amount | £3,608,961 (GBP) |
| Funding ID | EP/V038095/1 |
| Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 10/2021 |
| End | 09/2026 |
| Description | RAEng Chair in Emerging Technologies |
| Amount | £1,300,000 (GBP) |
| Organisation | Royal Academy of Engineering |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 03/2018 |
| End | 02/2028 |
| Description | UCB studentship: A combined omics and synthetic biology approach to enhanced CHO cell engineering' |
| Amount | £2,334 (GBP) |
| Organisation | UCB Pharma |
| Sector | Private |
| Country | United Kingdom |
| Start | 10/2018 |
| End | 09/2021 |
| Title | A degron controlled synthetic transcription factors have been developed |
| Description | The nuclease-deactivated variant of CRISPR-Cas9 proteins (dCas9) fused to heterologous transactivation domains can act as a potent guide RNA sequence-directed inducer or repressor of gene expression in mammalian cells. In such a system the long-term presence of a stable dCas9 effector can be a draw-back precluding the ability to switch rapidly between repressed and activated target gene expression states, imposing a static environment on the synthetic regulatory circuits in the cell. To address this issue we have generated a toolkit of conditionally degradable or stabilisable orthologous dCas9 or Cpf1 effector proteins, thus opening options for multidimensional control of functional activities through combinations of orthogonal, drug-tunable artificial transcription factors. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2017 |
| Provided To Others? | Yes |
| Impact | None yet it has only recently been developed and published. |
| URL | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5662744/ |
| Description | Engineering the Golgi apparatus in biologic producing CHO cell lines |
| Organisation | GlaxoSmithKline (GSK) |
| Department | Biopharm R&D |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | We are generating mutations in the vesicle trafficking machinery in the Golgi to enhance biologic production and control product quality, specifically glycosylation. We have also developed a computational modelling platform to link product quality (glycosylation) to Golgi homeostasis. |
| Collaborator Contribution | GSK are providing case studies to test our mutations and computational platform in their production lines with some of their target biologics. |
| Impact | A follow-on-funding application was approved and we are currently working on this. |
| Start Year | 2017 |