Tuesday, September 24, 2024 - Day One of Main Conference - ET (Eastern Time, GMT-05:00)
- Karthik Balakrishnan - CEO, Nodexus Inc
- Ron Weiss - Professor of Biological Engineering, MIT Synthetic Biology Lab
A team of BioPhorum members have developed and executed surveys focused on the activities and effort involved in a typical Cell Line Development (CLD) campaign. An average of 27 members from different companies that participate in the BioPhorum CLD working group answered surveys covering a standard CLD workflow from vector design to single cell clone selection. The surveys were very extensive, including a total of 341 questions split between antibody and complex molecule CLD processes. They provide a comprehensive industry perspective on the typical time and effort required to develop a CHO production cell line
- Thomas Kelly - Director, Cell Engineering & Analytical Sciences, Johnson & Johnson Innovative Medicine
Transposase/Transposon platforms have become increasingly common for the development of robust high-expressing CHO cell lines for protein therapeutic manufacturing. Notably, these techniques use a single transposase/transposon pair to enable such outcomes. ATUM, as part of the Leap In Transposase platform, has developed a number of mutually orthogonal transposase/transposon pairs that can be used to serially engineer CHO, and other, cell lines in a robust manner. Indeed, this engineering can be used to not only increase the expression of transgenes, as is the case for a mAb therapeutics, but also knock-down the expression of endogenous genes to affect cellular physiology and/or product quality attributes … or both. This talk will provide examples of such engineering including a case study wherein three orthogonal Leap In Transposase/Transposon systems were implemented for the creation of a mAb expressing cell line with specific product quality attributes.
- Oren Beske, Ph.D. - Amalgamator of Business and Biology, ATUM
- Ashish Saksule - Associate Director, Cell and Gene Therapy, Vertex Pharmaceuticals
- 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
- Nathalie Clement - Vice President of Vector Development, Siren Biotechnology, Inc.
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.
- Stacie Seidel - Senior Director Molecular/Viral Vector Biology, elevatebio
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.
- Luke McCarney - Filtration Engineer, Cytiva
In the biopharma industry, various techniques are utilized to enhance yield and quality of the target protein produced by stable cell pools and accelerate overall CLD timeline. In this presentation, we will show a case study of a method for minipool productivity enrichment via co-expression of the target protein with a fluorescent biosensor protein using an IRES, combined with state-of-the-art automation tools to allow productivity enhancement and reduce timeline for overall cell line development efforts.
- Jishna Ganguly - Expert Scientist, GSK
Cell Line Development plays a crucial role in establishing Master Cell Banks for clinical and commercial biomanufacturing. This involves creating subclones and undergoing multiple stages of rigorous assessment, leading to the selection of a final clone used for the project's entire duration. Decision-making in this process hinges on extensive datasets obtained from advanced analytical methods. The introduction of high-throughput platforms like the Berkeley Light Beacon and automated micro-bioreactor systems has resulted in generating vast datasets, which often consist of thousands of data points in each experiment. Moreover, the need to integrate process and performance data from various scales, including deep-well plates, shake flasks, and bioreactor processes, is essential for a thorough analysis. Collectively, these factors pose significant challenges in data processing and analysis, which are critical for informed decisionmaking in Cell Line Development. Here, we propose a holistic method for digitizing the entire cell line development and selection process. Our approach begins with implementing laboratory and data automation tools to streamline the generation and handling of raw data. We then establish automated data pipelines using the Databricks platform, enabling the integration of various data types and data of different scales into a specially designed database. This database comprehensively encompasses data on cell line creation, assessment, and selection. Additionally, we develop visualization dashboards linked in real-time to the database, significantly reducing time spent on data processing. Finally, we leverage this streamlined data to build predictive models using open-source Python machine-learning algorithms, enhancing the cell line selection process. Our proposed digital framework ensures a data-driven approach, optimizing the selection of highquality cell lines for clinical and commercial manufacturing purposes.
- Yi Li - Process Development Scientist, Amgen, Inc.
Great Bay Bio is a tech-bio company integrating big data analysis, AI, and automation into biologics development processing. We have developed an intelligent ecosystem covering biological drug discovery and CMC, including:
1. Antibody molecule development and optimization (AlfaDAX): within 1-2 weeks, the platform can develop and optimize antibody molecules with less wet-lab work in the aspects of humanization, affinity maturation, and developability assessment.
2. Site-specific integration technology for cell line development (AlfaCell): the stable monoclone with high titer (6-15 g/L) can be obtained within 1.5 months instead of the traditional process of 6 months, particularly solving two pain points of bispecific antibody development – lower titer and higher missing paring.
3. Non-screening cell culture media development (AlfaOPA): the customized cell culture media can be developed within 1 month only using 1 mL of the supernatant at the end of fed-batch processing.
The presentation will use obesity treatment target development, ACTRII, as a case study to show how the intelligent ecosystem developed a preclinical asset from drug discovery to CMC enabling within 7-9 months.
- Michael Chen, PhD - CEO & Co-founder, Great Bay Bio
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.
- Shiliang Hu, Ph.D. - Senior Scientist, Genomic Medicine Department, Alexion Pharmaceuticals
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 (PCLTM) 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.
- Sha Sha - Senior Scientist, Upstream Process Development, Ultragenyx
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.
- Nicky Young - Senior Director, Sterility Assurance, Teknova
CRISPR, a molecular technology developed by adapting a protective bacterial response to virus infection, provides previously unknown power and sophistication in editing genes in organisms from bacteria to man. Consequently, control over this technology, principally by patent protection, has engendered several waves of contested proceedings in the US and European patent offices, many of which are ongoing. Because there are multiple partied and multiple proceedings a definitive determination of who owns CRISPR patent rights and the accompanying royalties on licenses is something that will not be finally concluded for several years.
Nevertheless, or perhaps because of this situation the assignees of CRISPR inventors at several universities have engaged in a broad licensing regime covering application of the technology in medicine, agriculture, and other fields. This talk will walk through these regimes in light of the various academic and commercial interests and provide a glimpse at what may arise in future when these rights are more certainly established, as well as briefly touching on the existing companies that have already licensed CRISPR technology.
- Kevin E. Noonan, Ph.D. - Partner, McDonnell Boehnen Hulbert & Berghoff LLP
- Summarize key takeaways from discussions on accelerating CLD and FIH studies
- Encourage further exploration of innovative technologies and strategies for bioprocess development
- Highlight the importance of collaboration and shared learnings in advancing the field
- Thomas Kelly - Director, Cell Engineering & Analytical Sciences, Johnson & Johnson Innovative Medicine
- Charles Mitchell - Senior Process Scientist, Cell Culture, Visterra Inc
- Pitchai Sangan - Associate Director of Cell Line Development, Boston Institute of Biotechnology, LLC
- Nhu Nguyen - Scientist, Cell Line Development and Process Development, Aragen Bioscience Inc
- Nikki Nogal, PhD - Global Director, Technical and CMC, Lonza
- Grab a 'Cloud Spritz' (Aperol Spritz)
- Culture Bioscience's booth (328)
- Grab a SmartLabs 75 (French 75)
- SmartLabs Booth (1824)