Assessing Recombinant AAV Shedding After Cardiac Gene Therapy

Abstract

Gene therapy based on recombinant adeno-associated viral (rAAV) vectors has recently made significant progress as a clinical therapeutic. Unlike most traditional medications, gene therapy vectors can be biologically active when shed into the surrounding environment. Here we describe methods for collection and storage of multiple biological specimen samples and a PCR-based method for detection of shed adeno-associated viral (AAV) particles. We also describe a method for use of an infectious replication assay utilizing a cell line stably expressing AAV Rep and Cap genes and superinfection with adenovirus 5 to detect functionality in shed AAV particles.

Keywords: Adeno-associated viral vector; Biodistribution; Cardiac gene therapy; Infectious particle detection; Specimen samples; Vector shedding; qPCR analysis.