Improving the production and quality of lentivirus for the transduction of haematopoeitic stem cells via a perfusion culture process

Scalable and consistent production of LV is critical to ensure sufficient supply of high-quality vector for gene therapies and gene-modified cell therapies. The focus of the project is to optimise lentiviral vector (LV) production through the investigation of perfusion and the establishment of a high-throughput, scalable suspension platform for suspension-adapted HEK cells.

The process conditions established in the small scale stirred-tank bioreactor systems (100-250 mL) will be validated in larger-scale stirred-tank bioreactor system (up to 5L). The LV produced will be used to transduce the target cells of interest (hematopoietic stem cells - HSCs).

Specifically, this project will build on previous work undertaken at CSL developing a suspension-adapted HEK cell line and establish a robust perfusion process following large-scale DoE studies investigating the key process parameters.

Work will begin by investigating a range of platforms to establish a suitable high-throughput, small-scale suspension platform for the LV production studies. Once established, a perfusion process will be developed and the LV produced will be tested for functionality on the target haematopoietic stem cells.

This aligns to the EPSRC's Healthcare Technologies and Manufacturing the Future Themes, specifically the 'Developing Future Therapies' and '21st century products' Grand Challenges.

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.