This presentation will discuss: Heterogeneity of the starting leukapheresis poses challenges for successful manufacturing of autologous TCR T cells; cellular composition of the starting material impacts product characteristics; and optimization of starting material is critical to achieve adequate quantity and quality of the TCR T cell product.

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.

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.

Stirred suspension-based cell manufacturing can be used for scalable and controllable production of cell therapy products. We have developed custom reagents to modulate Notch signaling, which are compatible with suspension culture, and have implemented the production of pluripotent stem cell-derived CD8aß+ T cells in stirred tank bioreactors (STRs). STR-based culture enables process optimization and characterization using bioprocess solutions including automated feeding and in-process monitoring.

Human pluripotent stem cell (PSC)-derived natural killer (NK) cells combine the advantages of PSC and the safety profile of NK cells. At HebeCell we have developed our proprietary technology platform that is bioprocessing friendly and adaptable to GMP standards. Our feeder-free platform utilizes 3D spheroids to mimic the in vivo hematopoiesis to generate cytotoxic protoNK cells in bioreactors. Our platform offers a highly scalable approach for off-the-shelf cell therapies.

Analysis of the metabolome can be used for the evaluation of pluripotent and multipotent stem cell cultures, affording early detection of physiological changes indiscernible with traditional culture monitoring techniques. Finally, metabolomics can be used for the evaluation of the quality of stem cell differentiation in 2D and 3D biomaterial cultures, providing a sensitive and robust state-of-the-art technology, guaranteeing high-quality cell therapy manufacturing.

Currently available technologies for ex-vivo expansion of NK and CAR-NK cells using feeder cell expansion systems (e.g., K562 cells ) and cytokines (e.g., IL-2 in combination with IL-15) show several limitations telomere shortage, exhaustion, fratricidal killing, and regulatory concerns. Previous studies show that a human 721.221-mIL21 as a feeder cell can rapidly expand primary NK and CAR-NK cells. Based on this technology, we developed a non-feeder cell system to expand NK and CAR-NK cells. The results provided in this study show strong promise with the non-feeder cell expansion approach for functional NK and CAR-NK cells. 

Avenge Bio’s LOCOcyte platform consists of polymer encapsulated, allogeneic cells genetically engineered to produce immunomodulatory molecules for the treatment of previously intractable cancers. This presentation will highlight our approach to a successful technology transfer of our lead IL-2 program, AVB-001, into Phase I GMP manufacturing. In addition, we will discuss key considerations for future development and scale-up of this innovative cell therapy.

Explosive growth in the fields of informatics and cell and gene therapy (CGT) has occurred over the last four decades. Application of Informatics tools in CGT manufacturing can facilitate record keeping, quality management reviews and communications. An informatics backbone will be essential for deployment of disseminated manufacturing. While electronic records can completely replace paper and ink records, the potential benefits are much greater, including prospective quality management, error mitigation in manufacturing, release and administration, and reduction in redundancies. Implementation of informatics in CGT requires substantial planning, resources, and collaboration between CGT and informatics teams.

This presentation provides an overview of centralized and decentralized manufacturing models, pros/cons and suitable applications, approaches to overcoming challenges of decentralized manufacturing, and regulatory considerations.

Optimizing cell therapy manufacturing facilities is challenging due to small-scale cell therapy batch sizes. With batch sizes as small as one patient, manufacturers must scale out rather than up, duplicating every inefficiency in the process. This presentation explores four manufacturing approaches for small-scale cell therapies: dedicated rooms, chasing workstations, segregated unit operations, and process-in-a-box systems. Each approach has its challenges, and manufacturers must balance risk tolerance, capital spending, and growth expectations to achieve manufacturing goals. Case studies comparing facility designs that enable throughput will be presented and compared.