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.

The development of a robust manufacturing process for cell therapy products is a difficult strategy to navigate during the early stages. With the development of a solid manufacturing process comes the use of many raw materials to take your starting materials through to drug products. Common challenges and considerations will be discussed for assuring a successful manufacturing process that aligns with regulatory expectations.

This presentation will discuss various types of method qualifications, beginning with the exploratory work needed to refine validation strategies. It will cover the execution of qualification studies that validate and fine-tune these approaches, followed by the practical implementation of the methods. The discussion will conclude with best practices for accurately reporting the results, ensuring compliance and reliability in the application of these methods.

This presentation provides holistic approach to developing particulate control strategy for Cell Therapies with focus on particle characterization, visual inspection and manufacturing controls to understand sources and types of particulate matter, options for controls to reduce particle prevalence and consideration of patient safety. The application of these recommendations will enable development and enhancement of robust particle control strategies and provide more meaningful CMC engagement with health authorities with respect to particulate control and detection.

Automated blood culture and PCR-based methods have marked limitations for rapid sterility testing of CGT products. This presentation will discuss novel rapid sterility testing technologies, including a matrix-independent method capable of detecting microbial growth and identifying even slowest-growing bacteria and fungi in <72 hours, and most species within 24 hours, with high sensitivity (LOD <5 CFU) even in the presence of high cell concentrations.

Flow cytometry has become an essential tool in QC of advanced cell therapies while challenges in reproducibility remain. These challenges do not solely reside in laboratory-designed assays but instrument setup, qualification, and standardization still hold obstacles to overcome. We address a number of these challenges and obstacles within the context of the latest CLSI guidelines on Validation of Assays Performed by Flow Cytometry.

Analytics are essential parts of CGT CMC design. Analytic methods must demonstrate product sterility, identity, strength, and purity. Quality attributes for analytic methods require high specificity for the product, low limits of detection for impurities and contaminants, and high accuracy and precision. Validation of analytics must demonstrate consistent reproducibility. This requires sufficient repeated measures using consistent controls followed by statistical analysis to document confidence intervals for each analytic method.

The potency pathway for cell therapy includes developing assays early, tech transfer to QC, and lifecycle management. Regulatory agencies promote a risk-based approach for potency assay development. This presentation outlines key strategies for developing robust and accurate potency assays that meet regulatory standards, such as defining potential potency related attributes from the start and implementing strong QC measures to ensure consistent therapeutic performance.

Designing potency assays in early stages of product development poses multiple challenges. An unclear idea of the product’s mechanism of action, inevitable changes in manufacturing during development and scale-up, assay format, choice of product attribute for readout, choice of method for data interpretation—these hurdles will arise with program advancement. How should developers approach potency assay and assurance strategy development in a manner that will evolve with the product?

Cell-based assays are commonly used to characterize cell therapy products, typically as part of a potency matrix approach.  While these assays provide useful and often critical information about product functional attributes, they come with challenges including variability across operators, reagent lots, instruments, and laboratories. This presentation will focus on elements to consider when designing cell-based potency assays and relevant performance assessments, in order to support clinical product understanding.

This presentation will explore the critical role of characterization assays in the development of cell therapies. It will discuss how these assays enhance understanding of product attributes and production processes, leading to improved therapeutic efficacy and safety.

• Define benefits of integrative analysis of healthcare, digital health, and CMC data (improving efficacy through precision medicine, decreasing healthcare costs through companion diagnostics etc) 
• Outline risks to patients and donors (data reidentification, comprehensive LLMs, sales to 3rd parties etc) 
• Define stakeholder set (patient/donor advocacy organizations, pharma/biotech etc) 
• Discuss requirements for ethical LLMs/Agentic AI (confidentiality, accountability, integrity, compliance, competence etc)​

This presentation describes the journey of a stem-cell therapeutic & commercial development from Phase 3 and lessons learned. Particular challenges that exist within the stem cell therapeutics and regenerative medicine arena will be discussed. The application of QbD & DoE to both the process & assays was used to control the CPPs and CQAs for the product and how this approach aided successful PAI and BLA approval.

Analyzing surface proteins offers therapeutic insights, but their low abundance in cell proteomes complicates analysis. Developing workflows enriching for surface proteins would be a promising tool, especially in the field of cancer immunotherapy. CAR-T cells bridge the immune system with the tumor by engineering T-cells to recognize cancer antigens. Expression of the CAR is critical for therapeutic efficacy. Variations exist among similarly expressed CAR constructs, emphasizing the need to characterize associated proteins and identify post-translational modifications for improved outcomes. Our method involves immunoprecipitation of surface CAR followed by LC-MS analysis, providing valuable insights for enhancing therapeutic strategies.

Next-generation sequencing is revolutionizing the characterization of cell and gene therapies, addressing gaps in traditional methods. By employing bulk RNA-seq, single-cell RNA-seq, and genome sequencing, NGS provides detailed insights into critical quality attributes like genomic stability and cellular heterogeneity. This integration into quality-by-design frameworks optimizes manufacturing processes, ensuring product consistency and safety while accelerating development timelines. NGS stands as a pivotal tool in advancing therapeutic manufacturing: Enables detailed profiling of genomic stability and cellular heterogeneity; Optimizes manufacturing processes through data-driven insights; Integrates seamlessly into quality-by-design (QbD) strategies; Enhances product consistency and safety in therapeutic development; Accelerates timelines for bringing therapies to market.

Next-Generation Sequencing (NGS) as described in the ICH Q5A(R2). Virus Safety guideline ICH Q5A(R2) was published in November, 2023. Included are viral safety testing and evaluation of biotechnology products derived from well-characterized cell lines. In scope of this revision are genetically engineered viral vectors and viral vector-derived products. The talk will focus on the application of NGS to replace or supplement conventional virus assays for these new modalities.

The Advanced Virus Detection Technologies Working Group (AVDTWG) is a collaborative consortium focused on advancing next-generation sequencing (NGS) for detecting adventitious viruses in biologics. The group fosters scientific exchange among regulators, industry, CROs, and academia to address challenges in the NGS workflow including sample preparation, reference standards, bioinformatics, and follow-up strategies. Members of AVDTWG have developed collaborative studies for demonstrating the sensitivity of adventitious virus detection by NGS, and contributed to regulatory frameworks, including ICH Q5A(R2) and European Pharmacopoeia (Chapter 2.6.41). Together with IABS and PDA, AVDTWG supports knowledge sharing through conferences and webinars to advance viral safety in biologics.