Utilizing Multiplex ddPCR To Streamline Viral Gene Therapy Workflows

By Tara Quang, Senior Research Scientist, and Michael Baker, Senior Director, Viral Gene Therapy, FUJIFILM Diosynth Biotechnologies

Analytical technologies play a pivotal role in the development and successful commercialization of viral gene therapies. Among these, quantitative PCR (qPCR) has long been regarded as the "gold standard" for analysis of key quality attributes for viral vector characterization and batch release. For recombinant adeno-associated vector (AAV), this includes viral vector genome (VG) titer, purity, potency, host and process-related impurities, viral capsid characterization (i.e., the ratio of empty to full capsids), and the identification of replication-competent viruses.

While qPCR is a powerful molecular tool, it is limited to relative quantification, and the precision and accuracy of the results depend on how effectively the standard curve has been optimized. In the viral gene therapy space, there is a growing need for DNA quantification techniques that can more precisely quantify viral genome titers, which is essential for dosage determination in toxicology assessments and clinical applications1. Novel analytical technologies like Droplet Digital PCR (ddPCR) provide a more rapid and robust approach to nucleic acid quantification for therapeutic development that better align with the industry need for better sensitivity, accuracy, and precision.

In this article, explore the benefits of ddPCR over qPCR, key considerations for effective implementation of ddPCR, and how to leverage CDMO expertise.