Development of a scalable and consistent process for the production of lentivirus for gene-modified cell therapies

Scalable and consistent production of stable cell lines for lentiviral vector (LV) production is necessary to ensure sufficient supply of high-quality vector for gene therapies and gene-modified cell therapies. This requires an integrated approach with both biological engineering of the cell lines and bioprocess engineering of the upstream and downstream processing being considered in parallel.

The focus of the project is the development of scalable and consistent production of stable cell lines for LV production to ensure sufficient supply of high-quality vector for gene therapies and gene-modified cell therapies. Specifically, this will focus on the production of transient and stable cell lines for LV production to genetically modify clinically-relevant cells natural killer (NK) cells to express a chimeric antigen receptor (CAR).

The project objectives include:

Production of a LV coding for a marker gene, e.g. GFP, and a LV for anti-CD19 CAR via a stable, continuous LV small-scale production system
Identification and investigation of the key parameters for improved LV upstream processing using the iCELLis Nano via design of experiment studies including a comparison against other adherent technology systems.
Demonstration of end-to-end bioprocess for CAR-NK manufacture. This will be achieved via the transduction and expansion of the NK-cells with a chimeric antigen receptor (CAR) in the AllegroTM XRS system using LV vector produced in the iCELLis Nano.

Due to the multiple parameters and complexity of the process, an initial screening experiment will be established using a DoE approach to monitor virus recovery using simple, robust assays (p24 ELISA, qPCR and FACS-based infectivity assays). By mapping the full experimental space, we enable the use of Quality by Design in the final scalable process.

This aligns directly to the EPSRC Manufacturing the Future Theme, with a specific focus on manufacturing 21st century products. A major limitation at present for ATMP production is the lack of appropriate gene delivery vectors. This project seeks to address this critical bottleneck through scalable production of improved viral vectors.

The CDT has a proven track record of delivering impact from its research and training activities and this will continue in the new Centre. The main types of impact relate to: (i) provision of highly skilled EngD and sPhD graduates; (ii) generation of intellectual property (IP) in support of collaborating companies or for spin-out company creation; (iii) knowledge exchange to the wider bioprocess-using industries; (iv) benefits to patients in terms of new and more cost effective medicines, and (v) benefits to the wider society via involvement in public engagement activities and impacts on policy.

With regard to training, provision of future bioindustry leaders is the primary output of the CDT and some 96% of previous EngD graduates have progressed to relevant bioindustry careers. These highly skilled individuals help catalyse private sector innovation and biomanufacturing activity. This is of enormous importance to capitalise on emerging markets, such as Advanced Therapy Medicinal Products (ATMPs), and to create new jobs and a skilled labour force to underpin economic growth. The CDT will deliver new, flexible on-line training modules on complex biological products manufacture that will be made available to the wider bioprocessing community. It will also provide researchers with opportunities for international company placements and cross-cohort training between UCL and SSPC via a new annual Summer School and Conference.

In terms of IP generation, each industry-collaborative EngD project will have direct impact on the industry sponsor in terms of new technology generation and improvements to existing processes or procedures. Where substantial IP is generated in EngD or sPhD programmes, this has the potential to lead to spin-out company creation and job creation with wider economic benefit. CDT research has already led to creation of a number of successful spin-out companies and licensing agreements. Once arising IP is protected the existing UCL and NIBRT post-experience training programmes provide opportunities for wider industrial dissemination and impact of CDT research and training materials.

CDT projects will address production of new ATMPs or improvements to the manufacture of the next generation of complex biological products that will directly benefit healthcare providers and patients. Examples arising from previous EngD projects have included engineered enzymes for greener pharmaceutical synthesis, novel bioprocess operations to reduce biopharmaceutical manufacturing costs and the translation of early stem cell therapies into clinical trials. In each case the individual researchers have been important champions of knowledge exchange to their collaborating companies.

Finally, in terms of wider public engagement and society, the CDT has achieved substantial impact via involvement of staff and researchers in activities with schools (e.g. STEMnet), presentations at science fairs (Big Bang, Cheltenham), delivery of high profile public lectures (Wellcome Trust, Royal Institution) as well as TV and radio presentations. The next generation of CDT researchers will receive new training on the principles of Responsible Innovation (RI) that will be embedded in their research and help inform their public engagement activities and impact on policy.