TODAY’S CLINICAL LAB - OCTOBER 22 - 11:00AM ET
Nathalie Van den Berghe is a Senior Scientist at Trellis, a research group at the Catholic University of Leuven (KULeuven, Belgium) focused on AAV-based gene therapy. She completed her PhD in Pharmaceutical sciences at KULeuven where she developed immunoassays to evaluate drug and anti-drug antibody concentrations in patients. Currently, she is working as the analytics lead at Trellis and focusses on developing and optimizing assays, primarily for quantification of vector genome, capsid titers and the assessment of vector purity and integrity of AAV vectors for use as gene therapy products.
Accurate quantification of adeno-associated virus (AAV) genome copies in vector preparations is essential for optimization of the production and purification processes, preclinical studies and clinical dosage of AAV-based gene therapy products. Droplet digital (dd)PCR is one of the most widely used techniques for determining the AAV genome titer. Nevertheless, a consensus protocol for AAV viral genome titration is currently lacking. In this webinar, we discuss the challenges of viral genome titration and present an in-house developed and validated protocol for the quantification of AAV genomes in purified vector samples by ddPCR. Knowing the viral capsid titer, often determined by a separate method such as ELISA, the percentage of full AAV particles can be calculated. We discuss the use of the Vericheck ddPCR Empty-Full Capsid Kit which allows for the simultaneous determination of capsid and genome titer in one go, thus bypassing the need to perform separate assays to calculate empty-full ratios.
