Bioanalytical Methods to Study Biodistribution and Shedding of AAV ‐Based Gene Therapy Vectors

Biodistribution and shedding studies are instrumental for developing adeno-associated virus (AAV)-based gene therapies. The main objectives of these studies are to investigate where and in what form the vector is present within the body and whether measurable amounts are disseminated into the environment. In this chapter, we describe polymerase chain reaction (PCR)-based bioanalytical methods that measure AAV vector DNA in tissues and biological fluids. Considerations related to choosing real time vs. digital PCR platforms, principles of method development, key performance characteristics, and data reporting formats are presented. We also introduce and discuss a novel immunocapture-based PCR method for ultra-sensitive detection of intact AAV capsids.