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

This presentation will provide insight into the bit.bio team's recent work to scale-up from 2-D adherent culture to 3-D suspension culture of iPSC-derived cell therapies and commercial research products. The unique characteristics of bit.bio's foundational opti-ox technology allow for precise control of homogenous 3-D suspension cultures.

Here, we summarize the development and optimization of the scale down model for the BEAM-201 manufacturing process. The BEAM-201 clinical manufacturing process makes about 15x10⁹ edited allogeneic CAR-T cells to treat T-cell acute lymphoblastic leukemia (TALL). The large-scale (LS) process uses the cells from healthy donors that are selected based on the pre-defined donor screening tests. As part of the donor screening effort, the cell culture performance gets evaluated as small-scale in process development lab. Therefore, it is relevant to use a scale-down model (SDM) that performs comparably to the manufacturing process.

The BEAM-201 process consists of eight steps from isolating the cells to preparing the final drug product. Among these steps, the isolation was identified as the main source of variability across the two scales. By changing the isolation magnetic beads, columns, and reagents, we were able to obtain comparable process outcomes such as cell population doubling, purity, viability, and editing efficiency. We also report the addition of the TCRαβ depletion step to the SDM that could reliably predict the process performance at the manufacturing scale.

Cell therapies have emerged as a groundbreaking approach in the treatment of various diseases. However, the production process for these therapies is intricate and necessitates precise monitoring to ensure both safety and efficacy. Analytical assays that are commercially available have become essential tools in this process, seamlessly integrating into the workflows of researchers and manufacturers. These assays offer the capability to measure critical parameters such as safety, potency, and quality. During our discussion, we will explore the commonly used types of assays and demonstrate how these techniques can be applied to different stages of the production process. Additionally, we will introduce our latest solutions, including rapid sterility testing of samples, rapid mycoplasma detection, and assay kits that utilize digital PCR. Ultimately, this presentation aims to provide attendees with a comprehensive understanding of the significance of analytical kits in cell therapy production. Furthermore, we will highlight how leveraging these kits can improve the efficiency of manufacturing these life-changing therapies, reducing both the time and resources required. For Research Use Only. Not for use in diagnostic procedures.

• 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.

• Reducing wait times and logistical magnitude to reduce overhead costs
• Increasing accessibility, distributing to multiple locations closer to patient populations and remove transportation cost barriers
• How QC release testing and material kitting and management will be handled for POC manufacturing in this ecosystem.
• Establishing harmonized regulatory standards across different regions
• Implementing robust oversight mechanisms to ensure compliance with regulatory requirements and maintain product in a distributed manufacturing model.
• EMA vs FDA approach

Technology Transfer of Cell Therapy products into Good Manufacturing Practice (GMP) operations is a critical step in the commercialization of advanced therapeutic products. The process involves meticulous planning and coordination across multiple departments including development, Manufacturing Operations (including Quality Control), Quality Assurance and Regulatory-CMC. Herein we will discuss elements regarding facility set-up, documentation, process and analytical transfer strategies whilst considering project management do’s and don’ts form the viewpoint of ATMPs. Consideration will be also taken for both early and late phase products not only as a transferring site but as a CMO partner whilst considering various health authority requirements.

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.

METHONOVA™ is a methylcellulose designed for cell culture media. Plant-derived, xeno-free, METHONOVA™ is ideal for chemically defined media as a serum replacement or shear protectant for a variety of 2 and 3D culture processes. This talk will introduce METHONOVA™ features with several examples of its utility in cell culture processes.

Accurate assessment of empty and full capsids is pivotal for optimizing gene therapy vectors and ensuring their safety and efficacy. Existing methods for capsid analysis often need more time-consuming protocols and limited throughput, impeding progress in vector development. By developing a novel approach utilizing Droplet Digital PCR (ddPCR), rapid and robust evaluation of empty and full capsids is possible with high precision and accuracy. By quantifying proteins and viral genomes within droplets, the versatility of ddPCR technology can be applied across diverse biological samples, from crude lysates to purified samples. Data collected measuring empty-full capsid ratios aligns with the most stringent technologies with gold standard levels of precision. This innovative methodology holds immense promise for accelerating gene therapy research, ultimately advancing the development of safe and efficacious therapeutics.

The traditional drug discovery process is known for its lengthy timelines and high costs, relying on animal models that often don’t accurately predict human responses. DART for animal-free testing introduces a novel approach by employing human stem cells and advanced AI algorithms to develop testing models that closely mimic human physiology. This session will explore how DART’s integration of predictive analytics and stem cell technology could offer a more precise and potentially faster alternative to conventional methods. By better simulating human biological processes, DART may help identify promising drug candidates earlier in the development pipeline, reducing the risk of late-stage failures. Join us to explore how DART for animal-free testing can reshape the future of drug discovery, opening up new avenues for innovation while upholding ethical standards.

Based on client feedback, the most important challenge companies face when adopting CRISPR technology for commercial cell line engineering is the complex landscape of intellectual property (IP). Multiple ownership of CRISPR/Cas9 patents has created uncertainty regarding commercial freedom to operate (FTO). This presentation will showcase how Demeetra has combined its own Cas-CLOVER IP with the CVC, key holders of foundational CRISPR IP, clearing the path for our clients to obtain straightforward commercial licenses with undeniable FTO.

It is essential to select perfusion clones during cell line development (CLD) that thrive in the unique environment of intensified bioprocesses. Such clones have high specific productivity (qP), moderate growth rates, and high volumetric titer and yield. Traditional fed-batch CLD selects for clones that perform well in fed-batch, but this performance may not translate to a perfusion process. Here, we highlight the advantage of adopting a perfusion-specific CLD process to create more productive clones and provide an integrated solution for intensified processing using the CHOZN GS-/- cell line, a suite of best-in-class media, and the Mobius Breez Microbioreactor system.

Bruker's Beacon® platform offers comprehensive single-cell analysis by integrating light-directed Optoelectronic Positioning (OEP) and microfluidics, enabling deep interrogation of cell colonies through sophisticated assays and automated workflows. This accelerates cell line development by facilitating early evaluation of productivity, quality, and stability, streamlining clone selection in CLD campaigns.

• Grab a 'Cloud Spritz' (Aperol Spritz)
  • Culture Bioscience's booth (328)
• Grab a SmartLabs 75 (French 75)
  • SmartLabs Booth (1824)