This presentation will focus on the technical and manufacturing capabilities for cell therapy commercialization with discussion of CMC requirements, manufacturing launch readiness, and supply considerations. Specific considerations for a cell therapy and partnership with contract manufacturing organizations will also be discussed including lessons learned.

Increasing demand for recombinant adeno-associated virus (rAAV) based gene therapies necessitates increased manufacturing production. Transient transfection of mammalian cells remains the most commonly used method to produce clinical-grade rAAVs due to its ease of implementation. However, transient transfection processes are often characterized by sub-optimal yields and low fractions of filled-to-total capsids, both of which contribute to the high cost of goods of many rAAV-based gene therapies. Analysis of our previously developed mechanistic model for rAAV2/5 production suggested that the inadequate capsid filling is due to a temporal misalignment between viral DNA replication and capsid synthesis within the cells and the repression of later phase capsid formation by Rep proteins. We experimentally validated this prediction and showed that performing multiple, time-separated doses of plasmid increases the production of rAAV. In this study, we utilize the insights generated by our mechanistic model to develop an intensified process for rAAV production that combines continuous perfusion, high cell density transfection, and re-transfection which resulted in increased titer and plasmid efficiency. Our results establish a new paradigm for continuously manufacturing rAAV via transient transfection that improves productivity and reduces manufacturing costs.

AAV has emerged as a significant therapeutic modality in gene therapy. Challenges such as poor yield and variable product quality persist in the viral vector manufacturing space and we have addressed these problems using our cell engineering technology. Our cell engineering platform for improved adeno-associated virus (AAV) manufacturing addresses the critical challenges in gene therapy manufacturing, and presents an innovative modality for improving cells for production of viral vector therapeutics. Our platform, yielding cells with improved AAV production and CQAs, can significantly bolster the efficiency and cost-effectiveness of gene therapy manufacturing, and can accelerate current development timelines. We used a directed-evolution approach based on repeated cell fusions to shuffle the cell genome, and amplify chromosomes of HEK-293 host cells. Engineered clones enriched for mitochondria phenotypes were isolated, then used as transient-transfection hosts, and for creating stable packaging and producer cell lines. For generation of stable packaging and producer cell lines, we developed a novel inducible system that maximizes the capabilities of the inherent viral production machinery. Engineered HEK-293 clones grown in suspension culture exhibited up to 15-fold productivity improvement via triple transient transfection for AAV1, AAV2, AAV5, and AAV9 serotypes with capsid titers as high as 1017 viral particles/L (vp/L)—at least 10-fold higher than current industrial manufacturing processes. Selection for certain mitochondria phenotypes resulted in a 2-fold improvement in full-to-empty ratio—up to 55% full in crude supernatants. Finally, our engineered stable packaging and producer cell lines achieved capsid titers of up to 1016 vp/L. We demonstrated a multi-modal cell-engineering platform that has significantly improved yield and manufacturability for transient transfection and for stable packaging and producer cell line methods. We further propose a model regarding the role of mitochondria for enhancing capsid percent-full. Taken together, our disruptive platform technologies provide solutions for meeting current—and future—gene therapy manufacturing challenges.

• The development of first in class novel complexed miRNA-based cellular reprogramming therapeutics
• Exploring the use of miRNA for the treatment of Alzheimer’s Disease, Diabetes and Heart Disease

• Overcoming hurdles of viral vectors for Gene Therapy by harnessing the power of non-viral DNA-based platforms for Advanced Therapeutics
• Advancing Beyond Lipid Nanoparticles: Innovative Strategies in Delivery Vehicles Technology
• Exploring the use of DNA-based therapeutics for the treatment of diabetes, anemia and cancer in animal and human health.
• Case Reports

Automation plays a key role in improving the quality and lowering the cost of cell therapy products. As new or improved processing and testing technologies enter the market, it’s important for firms that build these products to design and provide both digital and physical integration touchpoints for their systems and consumables. This overview will discuss some of these important design considerations that will allow for more effective implementation of automation and make it easier to adopt new technologies.

Allogeneic cell therapies present a unique challenge for drug product process development by combining aspects of autologous cell therapies and conventional biologic products. As production scale increases to that of a commercial, off-the-shelf therapy, new challenges emerge for expanding fill finish and cryopreservation capacity. This presentation will discuss the developments and considerations that are necessary for pursuing large scale, allogeneic cell therapy manufacturing.

Ultragenyx is developing an investigational gene therapy, UX701, an adeno-associated viral vector-based gene therapy product, for the treatment of Wilson disease. UX701 is an investigational AAV9 gene therapy designed to deliver a modified form of the ATP7B gene. Ultragenyx has initiated Cyprus2+, a seamless Phase 1/2/3 study of a single intravenous infusion of UX701 in Wilson disease (NCT04884815). UX701 leverages Ultragenyx’s proprietary producer cell line platform, Pinnacle PCL™. This presentation reviews our first late-stage development activities for our upstream Pinnacle PCL™ platform process including parameter risk assessment, scale-down model qualification, and process characterization, to establish process controls for the upstream unit operations in our 2000L manufacturing process. The process characterization studies help us understand our parameter space and mitigate operational and quality risks which are critical for successful implementation of a robust commercial manufacturing process to provide consistent product quality throughout the product’s lifecycle.

Abstract TBC

Aurion Biotech is taking their off-the-shelf, allogeneic cell therapy (AURN001) from the lab to clinical trials with the goal of becoming the first mass-scale cell therapy to help restore vision to 17M+ patients around the world with corneal endothelial disease. Building a mass scale cell therapy means:

• Fulfilling a significant unmet patient need caused by the global shortage of donor corneas for transplant, in ways that are accessible and equitable
• Navigating the highs and lows of manufacturing and scaling an allogeneic cell therapy for clinical trials, successfully managing a CDMO partner and establishing a development process that can be translated into a large-scale commercial product.
• Creating a breakthrough product that will transform the physician experience – one that is less complex and less invasive than corneal transplant (the current standard of care) and minimally invasive for patients.

• How can early-stage/research professionals best prepare for GMP
• Critical aspects to keep in mind transitioning from early-stage development into more commercial manufacturing.
• Examples from experiences and lessons learnt
• Cross-team collaboration and pre-emptively preventing down the road bottlenecks

• How can early-stage/research professionals best prepare for GMP
• Critical aspects to keep in mind transitioning from early-stage development into more commercial manufacturing.
• Examples from experiences and lessons learnt
• Cross-team collaboration and pre-emptively preventing down the road bottlenecks