Meet our team at the ESGCT: Oct. 11-14, 2022 in Edinburgh, Scotland
The ESGCT conference will take place October 11-14 2022 at the Edinburgh International Convention Center in Edinburgh, Scotland.
Meet our team during the event:
Stop by our booth #20 to meet our team during the ESGCT conference and learn more about our extended services to bring your cell and gene therapies to life… From process development up to commercial launch, Yposkesi is the full-service CDMO of choice, working by your side, to meet your timelines and budget.
Cell & Gene therapy manufacturing – Current state and future developments, by Brian Mullan, PhD, CTO of Yposkesi.
Cell and gene therapy manufacturing has progressed rapidly since the development of early processes in 2000 – 2010, however, significant challenges remain. It could be argued that the current overall state as regards process and analytical development is something similar to the situation for monoclonal antibody development and manufacturing in the 1990s. Specific challenges include an under-developed technology offering, weak product understanding, and poor process economics (yields, cost of goods). An overview of the current state, and some key future trends will be presented. For AAV gene therapy manufacturing many manufacturing processes are now moving to larger bioreactor scales around 500 – 1000L, especially with the potential for commercialisation of therapies for non-rare diseases (e.g., muscular dystrophies) and manufacturing platforms using two chromatography steps are becoming a standard. For LVV, the situation is somewhat different, as the primary therapeutic category is for cell based immune-oncology therapy where the LVV is a critical staring material, and material quantity requirements are generally lower. To illustrate some of these points, data on bioreactor scale-up and also on transition from one-step to two-step chromatography processes for AAV will be presented for manufacturing process platforms used across client projects at Yposkesi. The cell and gene therapy industry clearly has an encouraging future ahead of it and will require continued evolution in process and analytical development to get there.
During the Developments in manufacturing and scale up track, on Thursday 13 October, at 15:30, Parallel 5b.
Improvement of adeno-associated viral vector Production using “HEK+” cells, deleted of SV40 large T antigen encoding sequences,
S. Charrier1, E. Triebe2, J. Cheuzeville2, L. Suarez1, O. Moses1, F. Amor3, N. Avenier1, C. Rousseaux1, M. Amendola3, P. Santambien2, B. Mullan1 and S. Martin2
1 Yposkesi, 2Genethon, 3Genethon, INTEGRARE, UMR_S951.
The majority of in vivo gene therapy clinical trials require the production of recombinant AAV vectors with high purity and potency. We have previously isolated a high producing clone derived from the human embryonic kidney (HEK) 293T cell line which grows in suspension in serum-free media, but the presence of the SV40 large T antigen may pose safety concerns. We have recently generated a new clonal cell line (called HEK+) derived from the HEK293T clone by removing the SV40 T antigen-encoding sequences via CRISPR-Cas9 genome editing. In this study, we have evaluated the performance of this cell line. Firstly, regardless of AAV serotypes (2, 6, 8 or 9), we have observed a two-fold increase in AAV productivity in comparison to the parental HEK293T clone in Shake Flasks (100mL) and also in 10L bioreactors, with titers higher than 1E11 VG/mL in the crude lysate. Secondly, we have evaluated the stability of this cell line for rAAV vector productivity up to 39 cell passages by measuring E1A and E1B mRNA expression. Finally, we have produced rAAV2/8 vector with a transgene of interest at the 50L bioreactor scale and confirm the ability to obtain high titers of purified rAAV vectors without SV40 T antigen expression.
Posters are located in Cromdale Hall on level -2 during the ESGCT conference, the presentation of our poster will be held by Sabine Charrier during the session on Thursday 13 October.