After assay validation, a control strategy should be implemented to ensure that the assay and control samples are behaving consistently with the validated state. Typically, trending analysis with statistical limits are utilized at this stage. This talk will present other statistical methods in addition to traditional control charting for assay monitoring. Establishing a baseline with inherent assay variability can be utilized to set acceptance criteria for assay monitoring.

Immunogenicity is a critical factor in biologics development, and proactive mitigation at the preclinical stage improves safety, efficacy, and reduces clinical attrition. We established an in vitro immunogenicity risk assessment platform within CMC to address risks associated with product-related attributes, including physical and chemical properties, and post-translational modifications. We will present case studies demonstrating the application of this platform to assess immunogenicity risk across different modalities.

Cell-based assays are essential for evaluating biological activity of biotherapeutics. The development of cell-based assays and the statistical processing of biological data present scientific and regulatory challenges. Key challenges include biological complexity, variability and reproducibility. To overcome these, choose suitable cell line to reflect MoA, implement rigorous experimental design and establish acceptance criteria.  Statistical considerations involve robust statistical methods and continuous monitoring of assay performance. Automation enhances consistency across studies.

Potency assays are critical for the development of biological drug products, and a potency assurance strategy is required. A case study will be presented on the development and qualification of bioassays for a bispecific antibody drug product. The potency assurance strategy for an early phase bispecific antibody drug product will also be presented.

Our study shows that post-translational modifications—specifically cysteinylation of a free thiol and sialylation of glycans—significantly affect the potency of a dual-receptor Fc-fusion therapeutic. Process-dependent differences in cysteinylation correlated with lower potency between batches, while chemical removal of sialic acids increased potency, highlighting the need for detailed product characterization to identify critical quality attributes during manufacturing optimization.

This presentation evaluates the robustness of an in-house BrdU-based T cell proliferation assay, analyzing the impact of experimental variables across more than 300 donor samples. Using bococizumab as a case study, we demonstrate that assay precision remains highly consistent when normalized to a negative control like bevacizumab. Additionally, we will explore how T cell responses correlate with specific HLA-DRB1 genotypes, revealing critical HLA-associated immunogenicity patterns relevant to therapeutic development.

Potency assays are essential for demonstrating the quality and biological activity of therapeutic products; however, even well-established methods can exhibit unacceptable failure rates or high variability. This talk presents case studies on the remediation of ELISA and cell-based potency assays for biologics, emphasizing root cause identification, targeted corrective actions, and restoration of method suitability for routine use. Through practical examples, the session presents a structured approach to improving assay reliability, reducing the risk of false out-of-specification results, and strengthening lifecycle assay management in alignment with current regulatory expectations.

This presentation describes the development of a confirmatory neutralizing antibody assay that combines immunoglobulin depletion from specimens with a hyaluronidase activity assay. Following method validation, an optimized three-tier NAb testing strategy was implemented: Screening of potentially positive samples, confirmatory testing of screen-positive samples, and titration of confirmed positive samples.

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.