QIAGEN GmbH Presentation: Analysis of DNA integrity and stability using digital PCR
Wednesday, September 4, 2024 10:30 AM - 10:45 AM
Room R18 – Global Showcase A
Global Showcase Presentation
Presenter:
- Francesca Di Pasquale, Director R&D - dPCR Applications Development, QIAGEN GmbH, Germany
Development of safe and effective cell and gene therapies is key to potentially treating a wide spectrum of diseases. Viral vectors have become powerful delivery vehicles for gene therapies. Adeno-associated virus (AAV) vectors have turned into primary modalities for efficient gene therapy applications due to their lack of pathogenicity and persistent transgene expression. Besides the requirement to quantify viral vector genome titers accurately and reproducibly, it is essential to determine the intactness of the viral vector genomes for a safe, stable and effective therapy. Of late, multiplex digital PCR (dPCR) has been adopted for genome integrity analyses. Digital PCR enables absolute quantification with unprecedented precision and a higher tolerance towards inhibitors without the need for standards. Here, we propose a rapid dPCR approach for characterizing genome integrity of in-process and purified AAV samples using the same primers and probes that have been optimized for vector genome titration. We show that up to five targets can be analyzed simultaneously, increasing precision and reproducibility of the analyses.
Session Objectives:
- Integrity status of AAV particles can be determined using digital PCR
- The more targets are being covered in a multiplex reaction the better the calculation of integrity
- Integrity calculation is robust over a broad dynamic range
