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A Novel, Perfusion-Based Lentiviral Vector Manufacturing Process Demonstrating Increased Productivity and Enhanced Purity

O Goodyear(1) C M Lamont(1) K E Pemberton(1) R Sanches(1) M D Kapanidou(1) G T Pamenter(1) J Wright(1) D C Farley(1) K A Mitrophanous(1) N Clarkson(1) J E Miskin(1) L A Davies(1) C Knevelman(1)

1:Oxford BioMedica

For over 25 years, Oxford Biomedica (OXB) has been a pioneer in the development of products and innovative technologies based on lentiviral vectors. To meet the forecast on vector demand for gene and cell therapies, OXB has recently introduced a next-generation transient lentiviral manufacturing process incorporating innovative process modifications which simultaneously enhance lentiviral process yields and improve vector quality attributes. This new process utilises perfusion technology to support vector production at higher cell densities than have been associated with traditional batch and fed batch production approaches for lentiviral vectors. This approach allows for significant process intensification without the need for increased bioreactor volume. Furthermore, using this technology, lentiviral particles secreted into the bioreactor, can be harvested using continuous perfusion which also allows stabilisation of generated vector at lower temperatures, leading to improved preservation of the final product.

Importantly, this process adopts a plug and play approach facilitating incorporation of small molecule enhancers, some of which have already been identified by OXB. For example, incorporation of U1 (RNA based enhancer) and I3A during vector production, using this new process, led to increased cell-specific productivity. Overall, process yield improvements from 2 to 10-fold have been demonstrated for a range of therapeutic lentiviral vectors with process scalability demonstrated in stirred tank bioreactors up to 200 L in GMP.

This new manufacturing platform offers significant benefits for clinical production of lentiviral vectors and will further enable OXB to support the continuing global demand for high quality gene and cell therapy products.

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