Implementing Design of Experiment (DOE) in ADA Optimization for Targeted Therapeutics

Purpose

In addition to Enzyme Replacement Therapies (ERTs), new modalities such as adeno-associated virus (AAV)-vectored gene therapies have shown great promise in the treatment of rare genetic diseases. The targeted activity of these therapeutics means that bioanalytical assays often need to be developed in unique matrices, such as cerebrospinal fluid (CSF), which may be limited in volume and contain interfering factors that complicate assay development. Therefore, approaches that shorten development and reduce matrix requirements are critical. Design of Experiment (DOE) is a statistical approach used to shorten the time involved in developing bioanalytical assays by testing the relationship between multiple factors simultaneously. Herein, we detail two case studies where DOE was implemented to streamline anti-drug antibody (ADA) assay development in CSF. The first case study details how DOE was used to design an ADA assay for AAV-based capsid with optimized sensitivity. The second case study applies DOE to developing a drug tolerant ADA assay for an ERT for a lysosomal storage disease with drug delivered directly to the central nervous system (CNS).