Tuesday, September 24, 2024 - Day One of Main ConferenceTuesday, 24 September 2024 - ET (Eastern Time, GMT-05:00)

• Roadmap on how to navigate the complex path of an AAV drug product clinical development from R&D to BLA
• Current trends on various Vector designs, Manufacturing platforms, PD and AD, assays for release and characterization from Phase I – BLA drug products
• Adapting to the evolving regulatory guidance for successful IND and BLA filing with the impact on CMC changes and comparability studies

Many cell & gene therapy companies struggle to find the right balance between screening a number of novel viral vector construct designs and moving quickly through development. Often the transition from R&D to Process development (PD) can be bumpy and may result in delayed timelines or, even worse, an underperforming candidate that does not meet critical quality metrics. Building in key manufacturability and quality metrics early on in R&D helps define a successful candidate. This involves employing advanced analytics and characterization testing at an earlier stage in R&D, including cell-based functionality, to help detect any red flags that may not have been found until later in development. Additionally, aligning protocols across R&D and PD early on helps simplify the coordination and allows for more streamlined interpretation of the data. The combination of these aligned workflows result in improved characterization and allows for more viral vector designs to be evaluated in parallel. The ability to screen and characterize more sequence designs helps to understand the impact of different designs improvements such as codon optimization, promoter variants and other sequence elements. We will demonstrate lentiviral vector and AAV workflows with representative data to help highlight a streamlined workflow for accelerated development.

As monoclonal antibody (mAb) platforms evolved, the roles for each tangential flow filtration (TFF) format settled into place. In upstream applications we take advantage of the low shear environment in hollow fibers to recirculate cells without impacting product quality. Flat sheet cassettes are typically used in downstream applications, as their higher turbulence enables faster processing in smaller footprints. However, platform processes for viral vector-based gene therapies are still in their infancy, and the roles for TFF technologies have yet to be defined. Interestingly, we have seen many examples of innovators using different technologies in the same application. These new modalities carry with them unique product and process requirements compared to mAb’s. For example, lower product stability may push us towards faster and gentler processing. And more challenging adventitious agent clearance may increase the need for closed processing. With these requirements in mind, is there an ‘optimal’ TFF option? Here we aim to address this question with a comparison of TFF technology formats: hollow fibers versus flat sheets. The work focuses on adeno-associated virus (AAV) and includes data from several common serotypes and multiple locations along the downstream process. We will share technical data comparing key performance outputs such as flux, purity, and yield. Included in that work we attempt to define flow and pressure limits based on product quality (i.e., shear sensitivity of the viral vector). Finally, our comparison will cover manufacturing considerations including footprint, process economy, scalability, and options for closed processing.

Plasmids play a crucial role in the production of Adeno-Associated Viruses (AAV) and significantly impact the quality of the final product. This talk will focus on three strategic plasmid optimization methods developed at Alexion Pharmaceuticals that have significantly enhanced both AAV yield and quality.

Recombinant adenovirus associated virus (rAAV) is the most widely used viral vector for in vivo gene therapy with over 200 ongoing clinical trials across the world. A manufacturing process that is sufficiently productive with the requisite product quality is critically important for commercialization success. The Ultragenyx production system for rAAVs is based on our proprietary Pinnacle producer cell line (PCL^TM) platform that leverages stably transfected cell lines with integrated AAV and helper genes, which are induced through infection with wild type Ad5. The production platform already generates a high rAAV yield with sufficiently high quality for various rare disease therapeutic targets. To better understand whether further improvements are possible at the cell line level, as well as to better understand the intracellular dynamics of rAAV assembly and secretion, we developed a mechanistic model, and used it to simulate rAAV expression and Ad5 replication. The model was trained with empirical scale-down production data. Sensitivity analysis and non-linear optimization were used to identify bottlenecks and provide insights into the critical cellular processes that contribute to high yield and high percentage of full rAAV particles. Analysis was conducted to assess the competition between rAAV and Ad5 production. Additional improvements to the Pinnacle PCL™ expression system were to identified based on model predictions which will provide a guide for our next-generation cell line engineering efforts.

Designing a production facility for gene therapy reagents requires meticulous planning to meet the quality and customization demands. Few facilities support the flexibility required nor meet the GMP standards for the small-scale manufacture of made-to-order products where sterility, process flow, and layout are critical. Learn how Teknova built their new, modular ISO 13485-certified facility to meet the rigorous demands of GMP-grade reagents for gene therapy development and commercialization.