Enhancing efficiency and quality of statistical estimation of immunogenicity assay cut points through standardization and automation

• A web-based customized assay statistical tool (CAST) is developed to automate statistical analysis for anti-drug antibody assays.
• CAST provides a tool to assess assay cut point at a click of the button during validation and in-study sample analysis.
• CAST takes into account multiple assay factors in cut point analysis for various experimental designs.
• Prediction limit approach, along with automated outlier detection and data transformation, is utilized to compute cut point.
• Equivalence evaluation is applied to assess the need for a group-specific cut point.

Biotherapeutics can elicit immune responses, which can alter the exposure, safety, and efficacy of the therapeutics. A well-designed and robust bioanalytical method is critical for the detection and characterization of relevant anti-drug antibody (ADA) and the success of an immunogenicity study. As a fundamental criterion in immunogenicity testing, assay cut points need to be statistically established with a risk-based approach to reduce subjectivity. This manuscript describes the development of a validated, web-based, multi-tier customized assay statistical tool (CAST) for assessing cut points of ADA assays. The tool provides an intuitive web interface that allows users to import experimental data generated from a standardized experimental design, select the assay factors, run the standardized analysis algorithms, and generate tables, figures, and listings (TFL). It allows bioanalytical scientists to perform complex statistical analysis at a click of the button to produce reliable assay parameters in support of immunogenicity studies.