BIOTECHNOLOGY
Droplet digital PCR (ddPCR) is
the currently used technique, offering a complete AAV viral titer workflow solution with an established track record, assay availability, GMP-compliant software for data analysis, etc. However, the workflow is laborious and limited by throughput.
THE MODERN APPROACH A recent development in the field has been the introduction of the QIAcuity nanoplate-based digital PCR system that closely resembles the easy sample handling of qPCR and generates the same level of accuracy and precision as the currently used ddPCR system but with much faster sample-to-result times (2h versus 7h) and higher overall throughput and scalability. Nanoplates are available with
approximately 26,000 partitions for applications where the target is rare and/ or diluted or 8,500 partitions in a 96-well format for high-throughput use with a concentrated template. Since AAV is usually produced with titers at or above 1x1,012 genome copies/ml, the nanoplate with 8,500 partitions is a good fit for this application. Using a single QIAcuity eight-plate
instrument, it’s possible to load 16 plates in an eight-hour shift for 1,536 wells. Te instruments can detect fluorescent signals in five different channels, allowing even more multiplexing, which may be valuable in further qualifying AAV genome integrity. Existing, well-characterised qPCR assays can typically be transferred to dPCR on the nanoplate-based system with very little
optimisation. Te system is designed to meet the biomanufacturing and quality control needs with software to assist with US FDA 21 CFR Part 11 compliance required in a GMP setting, audit trails, user management, installation qualification/operational qualification (IQ/OQ) service and services such as rapid on-site response time. Te nanoplate-based system has the potential to play a major role in the development of new manufacturing processes by keeping pace with the analytical
Developing safe and effective AAV gene therapies is critical
demands of iterative process refinements. By exploring integration with high-throughput liquid handlers and multi-condition bioreactors, biopharma companies can further shorten time-to-data for rAAV process development and accelerate the scale-up of gene therapy products.
Laura Mohr is with Qiagen.
www.qiagen.com
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