Cambridge Healthtech Instituteの第8回年次会議
Gene Therapy Manufacturing
2023年8月16 - 17日、EDT（米国東部標準時）
Registration and Morning Coffee7:30 am
OPTIMIZING PROCESS DEVELOPMENT
End-to-End Development of a Suspension Transient Transfection Gene Therapy Production Platform
Terrence Dobrowsky, PhD, Head, Gene Therapy Drug Substance, Biogen
Biogen is developing a suspension transient transfection production system to support programs within our Gene Therapy pipeline from preclinical to commercial manufacturing. This presentation will review successful development strategies for establishing critical biological raw materials, upstream systems, and downstream processes, as well as their scale-up to reliably generate high volumes of drug substance.
The Latest Advancements in Adeno-Associated Virus (AAV) Production and Purification at 1000L
Nitin Garg, Senior Director, Gene Therapy, Tech Operations, PTC Therapeutics
Despite the fact that the number of therapies utilizing rAAV has increased, there are still significant barriers to the use of AAV, such as low viral amounts and impractical purification processes.Here, we report the optimization of rAAV production in suspension HEK293 cells in SFM media via triple transfection approch. Titers >2e11 VG/ml were observed at harvest. Furthermore, optimization enabled several AAV serotypes & transgenes to exhibit a 3-4 fold titer increase. A scalable enrichment process was established to achieve >50% full capsids at DS. Development (10L), pilot (200L), and production (1000L) were used to determine the process's scalability and robustness.
Multi-Factor Optimization and Modeling Approach to Improve rAAV Titer for Early Clinical-Stage Gene Therapy Programs
Steven Wang, PhD, Senior Scientist, Upstream Process Development, Passage Bio
A scalable and cost-effective production process is crucial to making recombinant adeno associated viral vector (rAAV) gene therapy more widely accessible. In this study, multiple factors were optimized through a design of experiments (DOE) approach using flatware to improve the yield of early clinical stage programs using HEK293 adherent cell system. A model was established to systematically analyze the impact of each factor on titer response and the interaction of multiple factors using multivariate regression analysis. Conditions optimized in flatware were scaled into the iCellis Nano fixed bed bioreactor to test optimized conditions against a validated scale down model.
Coffee Break in the Exhibit Hall with Poster Viewing10:00 am
INCREASING TITERS, REDUCING TIMELINES
Comparison of Traditional and Novel AAV Production Processes
Matthew Roach, Associate Director, AAV Production, BridgeBio
There is an industry-wide need for improvement in upstream yield across adeno-associated virus production to meet eventual clinical and commercial need. Despite this, there is still a lack of information around the cellular processes and pathways involved in the manufacture of AAV. This presentation will describe our efforts to improve the yield of our upstream processes while simultaneously determining what cellular processes contribute to this increase in yield.
Process Development Strategies for Increasing the Genome Titer and Improving the Percentage of Full Capsids of AAV6
Bojiao Yin, PhD, Director, Vector Process Development & Manufacturing, ElevateBio
In this presentation, we will describe strategies applied in upstream and downstream process development to increase the vector genome (vg) titer and achieve high full particle enrichment of AAV6 particles using a two-chromatography step approach. With these improvements, the final AAV6 products showed higher concentration (>1E13 vg/ml) and better quality (70% full particle content) with reduced DNA and host cell protein (HCP) impurities.
Development and Optimization of a Modified-Batch Process for AAV Gene Therapy Using High-Throughput AMBR250-ATF Perfusion System
Wei Xue, PhD, Senior Scientist, Process Development, Ultragenyx Pharmaceutical
Accessibility and affordability remains one of the biggest challenges in AAV manufacturing, calling for a high-yielding, robust, scalable, and cost-efficient process. At Ultragenyx, we employ a HeLa producer cell/helper-virus based infection process for AAV production, which costs less and scales more readily than a transfection-based process. Furthermore, we developed a modified batch process with perfusion to boost productivity to > 5E11 GC/mL in up to 250L SUB.
Refreshment Break in the Exhibit Hall with Poster Viewing12:40 pm
INDUSTRIALIZING GENE THERAPIES
Industrializing Gene Therapies into Commercially-Viable Products
Johannes C.M. Van Der Loo, PhD, Director Clinical Vector Core, Perelman Center for Cellular & Molecular Therapeutics, Children's Hospital of Philadelphia
- Optimizing the process
- Cell engineering
- Cell line selection
- Reducing COGs
Refreshment Break in the Exhibit Hall with Poster Viewing3:00 pm
PLENARY KEYNOTE: LEADING TO TOMORROW'S ADVANCES
Current and Future Trends in Biomanufacturing of New Modalities
Konstantin B. Konstantinov, PhD, CTO, Codiak Biosciences
Using exosomes as an example, this presentation examines the current and future trends in biomanufacturing, and the technologies needed to manufacture emerging modalities at scale. Traditional biomanufacturing methods do not provide the industrialized, commercially scalable, highly efficient and reproducible manufacturing process essential for this new class of biotherapeutics- so we built it from the ground up.
The Digitalization of Biomanufacturing
Richard D. Braatz, PhD, Edwin R. Gilliland Professor, Chemical Engineering, Massachusetts Institute of Technology
A fully instrumented testbed is described for the end-to-end integrated and continuous manufacturing of monoclonal antibodies. The testbed consists of parallel bioreactors, simulated moving bed chromatography systems for capture and polishing, bespoke viral inactivation, and a MAST auto-sampling system. Experimental results are compared with a digital twin for continuous runs lasting 30 to 60 days each, which include variations in metabolites and glycosylation profiles in designed experiments. The increased consistency in the glycosylation profile of the monoclonal antibodies being produced is quantified when going from batch to semi-batch to perfusion mode, and when moving from start-up to quasi-steady conditions.
Networking Reception in the Exhibit Hall with Poster Viewing5:00 pm
Close of Day6:00 pm
Registration and Morning Coffee7:30 am
LENTIVIRUS PROCESS DEVELOPMENT AND QUALITY
Overcoming the Challenges of Biomanufacturing Lentiviral Vector
Martin Loignon, PhD, Team Leader, Cell Engineering, National Research Council Canada
The demand for lentiviral vectors (LVs) for R&D and engineering cell therapies stems from their efficacy to deliver genes into targeted cells. Current LVs' production bioprocesses vary widely, significantly impacting quantities, quality, and costs. We have used a holistic approach to address challenges of upstream and downstream bioprocesses to increase titers and recovery.
Considerations in Development of Lentiviral-Based in vivo Gene Therapy
Mukesh Mayani, PhD, Head of Process Development, Gene Therapy, National Resilience, Inc.
Lentiviral vectors (LVV) have demonstrated noteworthy clinical success in patients during ex vivo CAR T as well as stem cell therapy gene therapy (GT) applications. The third-gen SIN LVVs have shown improved safety profiles for a conceivable durable treatment of rare disease and cancer indications. However, it’s application as in vivo therapy option is severely limited due to manufacturing, safety, and quality challenges. In this presentation, we will highlight several development and manufacturing considerations for generation of LV vector suitable for in vivo GT use.
Coffee Break in the Exhibit Hall with Poster Viewing9:00 am
Subject matter experts sit down 1:1 with our moderator to discuss and share their personal bioprocessing experiences, insights, and advice. The real “pay it forward” atmosphere provides biopharma leaders with unique opportunities to leverage and apply their expertise to make better technology, process, and business decisions, and, ultimately, to accelerate success. Dedicated networking within the session allows all attendees to follow up and dive deeper into conversation.
OPTIMIZING PROCESS DEVELOPMENT
Advances in Process Development
Nick DiGioia, Manager, Process Development, LogicBio Therapeutics, Inc.
Implementation of a wide range of AAV capsid variants has provided a unique challenge to process development groups, as manufacturing attributes of the AAV differ drastically between serotypes. The Alexion team has developed a manufacturing process with the goal of improving the consistency of the productivity and the quality of AAV produced in the bioreactor, as well as providing flexibility in the purification process to handle performance differences between serotypes.
Scale-Up of Suspension 293T/17-Based Cells for GMP Manufacture of AAV
Bryan A. Piras, PhD, Director of Manufacturing, Children's GMP LLC, St. Jude Children's Research Hospital
This presentation will discuss scale-up of a suspension cell-based process for manufacture of AAV from 5 to 200 liters. Results, including cell growth, titers, and impurities, were consistent across 5 L and 200 L scales and will be presented along with several challenges related to scale-up and manufacture.
Sponsored Presentation (Opportunity Available)11:30 am
Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own12:00 pm
Refreshment Break in the Exhibit Hall & Last Chance for Poster Viewing12:30 pm
OPTIMIZING DOWNSTREAM PROCESSING FOR AAVs
Advanced AEX Platform for AAV Enrichment
Yonatan Abune, Principal Research Associate and Engineer, CMC, Vedere Bio II
In this investigation, we employed AEX chromatography to simultaneously enrich full AAV capsid and eliminate endotoxin from the final drug product. By optimizing the column conditions, we achieved a high level of AAV capsid purity, as well as a significant reduction in endotoxin levels. This strategy offers a promising solution for the production of safe and effective AAV-based therapeutics.
Lessons Learned: A Case Study in the Downstream Optimization of an AAV5 Production Process
Ashton Lavoie, PhD, Associate Director, Downstream Process Development, BridgeBio Gene Therapy
Manufacturing strategies and progress towards platform processes for adeno-associated virus (AAV) production have seen substantial advancement in support of the impressive clinical success for this modality. This presentation will provide a case study for the development of a robust, high yield downstream process for the production of AAV serotype 5. Key findings will be discussed in addition to pitfalls and challenges in this development work.
Using High-Throughput Techniques to Improve Downstream Process Development for AAV
Arjun Bhadouria, PhD, Scientist, Purification Process Development, Genomic Medicine Unit CMC, Sanofi
High throughput purification development has become an integral part of purification development by providing an order of magnitude reduction in material and time requirement. It can be especially valuable since material limitations are often a challenge for AAVs. An initial screening of several process operating parameters using small amounts of material can be performed to narrow down the development space to focus on and further optimize the process. Here we discuss the various high throughput approaches utilized in Sanofi for developing several downstream unit operations including clarification, affinity chromatography, polishing AEX chromatography, etc.
Networking Refreshment Break2:40 pm
Studying AAV Capsid Aggregation in Complex Matrix of Clarified Lysate
Yulia Ivanova, PhD, Principal Scientist, Bioprocess R&D, Pfizer Inc.
Recovery out of harvest is the least understood step in downstream purification of AAV vectors. Complexity of lysed cell culture coupled to relatively low protein concentration of AAV product makes it very difficult to analytically investigate this process space. Here we evaluate the ability of dynamic light scattering (DLS) to serve as analytical characterization tool that would allow to investigate AAV capsid aggregation in a complex matrix of clarified harvest.
Plasmid Purification Process Development for Gene Therapy Applications
Close of Summit4:25 pm