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.

The manufacturing and release of cellular therapy products (CTPs) requires high quality, robust, and validated analytical methods. Here I will describe recent efforts in standards development and public-private partnerships to support the development of critical analytical methods used in advanced therapies. A key aspect of analytical development for these new class of products is the need for a fit-for-purpose approach when identifying CQAs and designing appropriate analytical methods. Cell viability is a fundamental measurement for cell-based therapies which also requires a fit-for-purpose approach. A framework for establishing fit-for-purpose cell viability assays will be described as well as ongoing standards efforts.

In this presentation we will discuss real world examples of how to build quality into the CMC and how real world deviations and incidents have led to a better quality program. Examples will include improvements to change control, training, operations, and testing.

In this presentation, we will look at some of the key requirements in testing and characterization requirements in cellular therapies. We will also discuss CMC considerations for product characterization and safety testing in early phase clinical trials of cellular therapy products.

The development of cell therapies, such as engineered Chimeric Antigen Receptor (CAR) T cells has great promise for patients. As the field continues to better understand the biological outcomes and clinical science, many new therapies are being pursued targeting different potential antigens or cancer indications. In this presentation, we will discuss early development considerations that can set up sponsors for success as their programs mature into late-stage. The decisions regarding process equipment, growth media, cytokines, and analytical methods (e.g., release vs. characterization) can be assessed through a staged approach and companies can benefit from early definition of a target product profile and analytical control strategy.

We explore how an objective technology transfer approach should be applied for cell therapy products and provide detail of the mechanisms and tools which should be used in order to make sure that the transition from development to cGMP is correctly achieved.

Tigen Pharma and CHUV in Lausanne, Switzerland, have formed a unique and close collaboration in development of tumor-infiltrating lymphocytes for treatment of solid tumors. The deep scientific knowledge of the Coukos and Harari teams at CHUV and the industrial expertise in Tigen accelerates the path of product to patient. This talk presents examples of this collaboration for progress in manufacturing and analytics for TIL in future clinical trials.

Base editing is a next-generation approach to gene editing with single base precision. I will review academic pioneering work on base editing based on existing publications. I will focus on analytical development, adding value to essential steps from bench to bedside, in my presentation. Two case studies will be discussed: potency assay development and how analytical assays contribute to cell therapy drug development using a quality-by-design (QbD) approach.

• Trends in CMC
• ​Tech transfer
• Manufacturing

• Importance of the cell therapy potency assay
• Potency assay development challenges
• Bridging the gap between R&D and Manufacturing
• The evolving regulatory landscape of potency assays​

As we have gained experience with analytical methods, we have learned more about the pain-points associated with our assays. With renewed focus on efficiently bringing our pipeline into the clinic, we have taken this opportunity to redevelop how we measure lentiviral vector potency. This talk will discuss our new methodology, including approaches to improve assay throughput, improve consistency of the readouts, and reduce assay complexity.

In the C&GT field, process-related DNA impurity levels are a safety focal point. Here, we provide the statistical analysis results based on several DoE studies  or reduction of hcDNA and pDNA levels in our Lentiviral Vector (LVV) platform process. Focusing on several critical parameters including nuclease concentration, supplement concentration, incubation time, and incubation conditions (ph/temperature), this data shows the optimal factors and ranges in the nuclease digestion step. 

Rare cell analysis on large numbers of cells is important in characterization of cell products as unwanted effects can be caused by just a few cells. Conventional flow cytometers are designed to accommodate assay development, allowing operators to fit instrument settings to their specific needs. The 'open' design of this instrument brings a significant level of complexity to flow cytometry and challenges reproducibility in manufacturing processes of therapeutic cells.

Live-cell imaging can provide quantitative dynamic and spatial characteristics of iPSCs in culture. Our research is geared toward developing systematic workflows quantifying the effects of culture and other handling conditions on iPS cell characteristics such as rates of mitosis, changes in morphology, and expression of relevant transcription factors to predict and evaluate the effects of manufacturing conditions and other perturbations on the state of the population. 

CAR-T cells are engineered T cells expressing a chimeric antigen receptor (CAR) on the cellular surface. CARs allow the T cell to engage an antigen on tumor cells and activate downstream signaling that leads to destruction of the tumor cell. Surface expression of the CAR is critical for therapeutic efficacy, but often differences in efficacy are observed between constructs that show similar surface expression. We developed a workflow using immunoprecipitation of the CAR from the cell surface to characterize post-translational modifications by LC-MS (IP-MS) to gain insights on differences in efficacy.

Accurate identification of recombinant plasmid and lentiviral vector sequences is important for raw material release in cell therapy. Our team has made significant advancements in analytical method development for sequence identification and variant detection through NGS platform. These methods have also proven invaluable for characterizing cell therapy products for CAR-T. In parallel, we have also developed an innovative multi-product sequencing approach that has enabled the identification of impurities across in cell therapy products. This includes the ability to differentiate single gene from multi-cistronic cell products. Recent advancements in method development for CRISPR on/off target detection using NGS and ddPCR platforms will also be highlighted.

The nature of cell therapy products means they cannot be terminally sterilised or even sterile-filtered; this puts a greater onus on other aspects of sterility control. This talk will discuss how a holistic approach is needed to ensure safety of these products, including facilities, testing and methods of sterilisation.