Viral vector-based gene therapies continue to grow with several products now approved and a flowing pipeline of candidates seeking approval in the coming years. To date, the most commonly used vectors are adeno-associated virus (AAV) based. The FDA does not currently provide guidance on how to approach immunogenicity assay development and validation when using emerging or existing technologies - a requirement to determine exposure in the preclinical and clinical phases of drug development. As a result, there is an increased need for scientific expertise to develop bioanalytical strategies utilizing new technologies as well as existing platforms within the regulated bioanalysis space to assess the safety of new AAV products. In this eChalk Talk, we will discuss considerations for AAV-based gene therapy programs, and best approaches and strategies for development and validation of assays to support regulated bioanalysis.