KCAS will present a poster at the American Association of Pharmaceutical Scientists (AAPS) National Biotechnology Conference (NBC) in Philadelphia later this month. The poster will compare droplet digital PCR (ddPCR) and quantitative PCR (qPCR) sample handling protocols for detecting and quantifying adeno-associated virus (AAV) vectors in any sample.
What is the poster featuring?
Carrie Vyhlidal, Associate Director of Cell & Gene Therapy at KCAS, submitted the poster on behalf of KCAS and will be in Philadelphia to present the poster during this year’s AAPS NBC. “In this work, we compare both qPCR and ddPCR to accurately quantify genome copy numbers of an AAV2 vector, similar to those that are used to deliver a gene therapy,” said Vyhlidal. “Publications in the literature describe different methods of how the sample has to be treated to get an accurate quantification of the genome copy numbers. Some of them involve DNAse treatment, to remove any unpackaged DNA that might be in the sample that will interfere with the detection of encapsidated DNA. Another critical step is breaking open the viral capsid before PCR to release the DNA that the virus is carrying. Some methods use proteinase K treatment to break down the viral capsid while other methods just use heat treatment to open up those capsids. We wanted to compare some of these methods and their performance both in qPCR and ddPCR to see if we could determine an optimal method, at least in our hands, to quantify genome copy numbers.”
AAV vectors are commonly used as methods to deliver gene therapies, and qPCR and ddPCR are being used more frequently to quantify genome copy numbers of AAV vectors in pre-clinical and clinical testing during product development due to ease of implementation, high sensitivity and accuracy, high throughput. However, there are very few regulatory requirements for these PCR-based methods, which has raised questions about what’s appropriate for these platforms with respect to method development and validation for biodistribution and shedding studies. The team at KCAS conducting this research includes Dominic Warrino, Senior Director of Scientific Services; Matt Pennington, a Senior Scientist; and analysts Brianna Longacre and Amanda Souaysene.
Who should attend this poster presentation?
The poster will be of interest to anyone who needs to quantify any adeno-associated virus viral vector in a gene therapy, whether it’s preclinically in animal models or even in a clinical study. Although the team has focused on AAV2, they believe that these methods could be expanded to other AAV serotypes.
The poster presentation is an opportunity for those in the field to learn more about the comparison of ddPCR and qPCR and the sample handling protocols for detecting and quantifying AAV vectors. If you’re attending AAPS NBC, make sure to stop by KCAS’ poster presentation to learn more about their research.