Bioassays are key to evaluate potency and safety of genome editing therapies and ensure they accurately target and modify the genome with minimal off-target effects. We developed a clinical bioassay using exosomal RNA/protein as non-invasive pharmacodynamic biomarkers to assess treatment response and proof of drug mechanism. We also developed methods to assess germline editing with a sensitive assay to provide reassurance for the absence of editing in human sperms.

Statistical Process Control techniques are often used to monitor bioassays. These techniques typically assume that values of the monitored metric (often model parameter) are independent. However, dependence structures often exist in bioassay data (i.e., replicate observations of a metric within an assay tend to be more similar to one another than between assays). In this talk, we explore various strategies for the valid application of SPC techniques in bioassay monitoring.

Confidence intervals for relative potencies are a major target of the evaluation of biological assays. This Monte-Carlo study compares the different compendial methods to calculate confidence intervals for relative potencies (reportable values) for biological assays.

The use of cells in bioprocessing and therapeutic products requires high-quality, robust, and validated analytical methods for cell characterization. Here I will describe recent developments in analytical method standardization, including NIST laboratory programs that support documentary standard and reference material development, and public-private partnerships to support the development of critical analytical methods. A key aspect of analytical development is the need for a fit-for-purpose approach.

The rise of new non-protein drug modalities, like plasmids, is thrilling. However, it also brings challenges in creating appropriate bioassays and fulfilling validation standards. This case study details the creation of a cell-based assay for potency testing of a plasmid active pharmaceutical ingredient (API), highlighting the difficulties faced throughout the development and validation process.

Effective mitigation of the immunogenicity of biologics necessitates a comprehensive understanding of its underlying mechanisms and multiple risk factors, enabling the identification of potential intervention points. Numerous academic and industrial organizations, along with US- and Europe-based scientific societies, have formed collaborations to address these issues. This presentation will provide an overview of ongoing and emerging initiatives within various frameworks.

A key element of drug safety and efficacy is developing therapeutic protein with extended half-life to deliver desired therapeutic outcomes. Historically, attaching Fc region (Fc-fusion protein) to the drug molecule has been implemented for recycling drug circulation in the serum. Another approach for pharmacokinetic (PK) enhancement is connecting drug molecule to the human serum albumin (HSA). We propose to compare and contrast the analytical control strategy for the two methodologies.

• In vitro assays for characterization
• Using automation in bioassays: When, where, and how to integrate
• New modalities including cell and gene therapies, immunotherapies, oncolytic virus therapies, and antibodies
• Potency assay development
• New bioassay technologies to increase speed and sensitivity

• Designing assays to fit into a clinical workflow
• Bioassays to understand a drug's Mechanism-of-Action (MoA)
• Designing bioassays for biologics with multiple Mechanism-of-Action (MoA)
• Robotization of cell based assays: When to use it and when not to use it
• Reference Standards for innovative drugs
  

The development of innovative biopharmaceuticals requires a comprehensive understanding of their mechanisms of action (MOA) to ensure their effectiveness and safety. Understanding the MoA of these agents is crucial for optimizing their therapeutic use and predicting patient response. This presentation focuses on our efforts to develop and implement cell-based assays using reporter cell lines that provide deeper insights into the MoA of biotherapeutic agents. We will discuss the rationale for choosing an analytical strategy: specific analyses and the methodology used. This approach accelerates the drug discovery process, ultimately contributing to the development of more effective and safer biopharmaceuticals.

Various autoimmune diseases are driven or worsened by autoreactive T cells attacking organs, tissues, or transplants. We created an assay system using patient-derived PBMCs and artificial antigen presentation cells to model T cell activation against a particular tissue. We demonstrated targeting antibodies fused to Ig-like domains from various immune checkpoints can block T cell activation, demonstrating the promise of selectively potent and specific T cell modulation biotherapeutics.

The multiplex opsonophagocytic assay, with colony counting as the assay endpoint, is critical for assessing pneumococcal vaccine immunogenicity. Current counting methods are time-consuming and prone to variability. We developed a deep-learning pipeline for automated colony segmentation and counting, which achieves superior accuracy and minimizes human intervention. This scalable solution is targeted at enhancing the efficiency of the assay workflow by 30% to support faster and data-driven development of pneumococcal vaccines.

The development of mRNA-based medicines requires robust potency assays to assess both translatability and biological activity. These assays can be either cell-free or cell-based assays; each provide a different assessment but are complementary to each other. As such, we will discuss various strategies for assessing mRNA DS and DP potency evaluation and the advantages and disadvantages between cell-free and cell-based assays.

Accurate potency monitoring is essential in drug development to ensure efficacy and safety at every stage. This presentation explores the application of T cell activation reporter gene assays as key tools for potency evaluation. We advocate exclusively using these assays, which leverage genetically encoded systems in T cells to measure immunological activity and offer a more accurate reflection of mechanism of action than single target binding assays. Join us to learn how T cell reporter assays are transforming potency monitoring and advancing drug development strategies.

Product potency is a critical quality attribute but not all assays designed to measure it will correlate with clinical outcomes.  Regulatory guidance indicates that potency assurance for a product should include acceptance criteria throughout development and result in rejection of sub-potent lots. To establish criticality, product batches with different potencies need to be evaluated clinically for their impact on effectiveness and safety. Phase appropriateness will be discussed in this context.