- Onboarding new trials at Dana Farber Cancer Institute 

- Set up study specific timelines

- Use change data to set up clear expectations with internal and/or external sponsors

Cellular therapy products (CTPs) require high quality, robust, and validated analytical methods. In recent years, several NIST-led ISO standards have been developed that address common testing needs for CTPs including cell characterization and count, and current efforts aim to develop a cell viability standard. Here, we describe the recently published and upcoming standards and the cell-counting COMET application, and give practical examples for the development of fit-for-purpose analytical methods.

Flow cytometry assays have been used to measure critical quality attributes, including viability, identity, purity, and potency of cellular therapeutic products. However, the lack of result comparability remains a significant challenge. NIST launched Flow Cytometry Standards Consortium by providing metrology and standards development expertise to work with the consortium members and stakeholders for developing measurement solutions and standards needed to accelerate translation, manufacturing, and approval of cell and gene therapies.

The implementation of Process Analytical Technologies (PAT) for cell therapies is needed to further process product understanding; however, few technologies enable real-time, continuous monitoring of product quality attributes. We utilize in-line holographic imaging through experimental, computational vision and machine learning techniques to predict multiple CAR T cell phenotypic attributes within the expected measurement variability of offline analytical methods. This approach enables phenotype monitoring across a variety of processing modalities. The implementation of PAT in cell therapy monitoring of CQAs would enable end-to-end data collection and novel control strategy development, providing the tools to create consistent and robust products.

Flow cytometry is a crucial analytical tool for in-process monitoring, release of cell therapy drug products, and clinical monitoring of patients. Despite its prevalent use, its complex design and high-dimensional capabilities require substantial consideration during development of flow-based analytical procedures and also for subsequent routine execution in GMP settings. Herein, we will discuss the current flow cytometry-related guidelines, challenges, and approaches to overcome them.

USP is continuing to develop reference standards, informational chapters, and compendial analytical methods to safeguard raw, starting, and ancillary materials for cell therapies. USP’s standards give best practice guidance to developers and manufacturers, simplify risk assessments, accelerate analytical development, and support raw material qualification and release. This presentation will describe existing standards and USP's recent development related to plasmid DNA and rapid microbial methods.

Compared to other medicinal products, cell therapy products (including gene-modified) tend to use a lot of biological raw materials. These can be human, animal, microbial, or even plant-derived. Without understanding how these materials are made, it is not possible to ensure their adventitious agent risks are addressed. Using real examples, this talk will discuss the principles and how to assess and mitigate the identified risks.

The regulatory expectations for potency of CGTs remain critical to consider during drug development. A review of the recent FDA guidance will be discussed, along with considerations to reduce risk with respect to potency assurance. Significant challenges will be encountered during potency assay development. While the assessment of potency may change over time, it is imperative that potency assays are developed incrementally and in parallel with clinical development activities.

Developing potency assays and demonstrating that they measure appropriate biological activities has long been a significant challenge, particularly with the evolving regulatory landscape. But what if you are developing a pioneering new therapy with a multimodal mechanism-of-action? This presentation will showcase the challenges of developing a potency assay matrix for an engineered macrophage therapy and will consider how these assays support future commercial manufacturing strategies.

A well-designed potency assay ensures lot-to-lot consistency of cellular drug products and contributes to the reliability of the drug development process. Identifying and understanding the correlation between multiple cell product potency critical attributes and release assays designed to monitor them at an early stage has many benefits. Early commitment to these potency assurance strategies reduces the risk of poor potency assay design, accelerates product development, and avoids regulatory hold-ups.

The B cell is a powerful cell that produces thousands of proteins per cell per second at constant levels, over decades. Precision genome editing can now be used to engineer B Cells that produce therapeutic proteins of interest, driving a new class of cellular medicines – Engineered B Cell Medicines (BCMs) – with the potential to be durable, allogeneic, redosable and administered without pre-conditioning. The promise of BCMs could transform therapeutic biologics with broad application — across protein classes, patient populations and therapeutic areas. Analytical strategies will be discussed. 

Neurona Therapeutics is a clinical-stage biotherapeutics company developing an allogeneic GABAergic inhibitory interneuron cell therapy candidate (NRTX-1001) for drug-resistant MTLE. NRTX-1001 clinical product is manufactured at Neurona’s cGMP facility, cryopreserved, and delivered to the clinic for MRI-guided deposition into the seizure-onset region of the temporal lobe. Analytical development that underpins manufacturing of NRTX-1001, the scientific background of the therapy and early first-in-human clinical data from the ongoing open-label Phase 1/2 study (NCT05135091) will be discussed.

The ability to quantitatively image induced pluripotent stem cells (iPSC) to monitor their dynamic and spatial behavior and state of pluripotency in a non-invasive manner is important for establishing better metrics for pluripotency and to assure consistency and efficiency in iPSC manufacturing.

This session is about nurturing the seeds of a good CGT concept to a healthy plant ready to bear the fruit of early phase clinical study. Seeds and young plants need fertilizer (funding), planting (tech transfer), and proper gardening tools. This talk will address effective communication between research laboratories (seed planters) and early phase CDMOs (plant tenders). Through dialog, the two sides of the tech transfer may grow research lab methods, applying new tools and a CMC focus, to arrive at manufacturing methods designed to continue growth through early phase clinical trial through to full commercialization.

Autologous therapy is not for marketing but only for the single patient using patient cells. We need new concepts to regulate autologous cell therapy. Autologous cell therapy is not for commercial distribution. Should CAR T therapy be regulated as a commercial product? DC therapy using patient idiotype antigen studies showed survival difference but no followers. New regulations started covering advanced therapy and US FDA approved autologous therapy for multiple myeloma last year. Japan's regulation has two pass ways: commercial product license application via PMDA or advanced therapy application via MOHLW.

Proteins are subjected to a myriad of modifications (PTMs), such as spontaneous degradation (e.g., deamidation and hydrolysis), oxidation, and crosslinking (e.g., via histidine). Moreover, for complex systems such as gene and cell therapies, higher order structures (HOS) also play critical roles. In this talk, their analyses and contributing factors, such as reactive metabolites (e.g., methylglyoxal, MGO) and cell culture changes, will be discussed, as well as the potential effects (e.g., immunogenicity and off-target binding). One pertinent issue is the chemical and physical degradation during cryopreservation of cells, and potential remediations are proposed.

This presentation will discuss optimizing containers, volumes, and formulations-dependent freezing profile to maximize yield on cryopreservation; best practices on the cell bioprocess from freezing through thawing, to maintain consistent cell viability; post-thaw cell characterization (ex: apoptotic markers) for process development and to reduce cell death; novel methods on cryopreservation for future cell therapies.