Optimizing AAV purification processes with mass photometry
Analytical techniques and challenges in AAV purification protocols
Adeno-associated viruses (AAVs) are a widely used vector for recombinant genome delivery into cells, and promising vehicles for novel gene therapies. The production AAV-based gene therapies require optimized purification processes that maximize the yield of loaded AAV capsids. ​
Purification represents a major challenge for AAV process development, and samples need to be characterized at several stages of the AAV purification protocol. Current gold standard techniques for AAV analytics such as analytical ultracentrifugation (AUC), electron microscopy (EM) or qPCR/ELISA present drawbacks such as long turnaround times, high sample consumption and need for highly specialized staff or outsourcing. ​
Mass photometry can support AAV purification protocols
Mass photometry is a powerful tool for AAV process development. Mass photometers like the SamuxMP and SamuxMP Auto are tailored for AAV analytics, providing comparable data to gold standard methodologies for AAV analytics with several advantages:
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Accurate, single-particle characterization within the relevant mass range (0.5-6 MDa)
Detects impurities, heterogeneous capsid loading and aggregation in <5 minutes
Requires as little as 5-10 µL of sample with a concentration of 10^11 particles/mL
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These strengths make mass photometry an ideal technique to support development and optimization of AAV purification protocols. This single-particle method detects even small quantities of impurities, and low turnaround times allows for multiple measurements at different stages of the AAV purification protocol. Finally, low sample requirements make mass photometry practical to implement in early process development, where AAV samples are small and valuable.
How to streamline AAV purification protocols with mass photometry
In this application note you will learn how mass photometry can be used to evaluate a sample at different stages during AAV purification, supporting AAV process development. You will see mass photometry analyses of samples collected at various stages during downstream AAV purification, including data on empty/full AAV ratios as well as on the presence of other impurities – such as cellular debris, partially filled and overfilled capsids, and aggregates. The data shown was obtained with the SamuxMP. Download the application note to learn the details.
Additional resources​
WEBINAR​
Streamlining AAV characterization with automated mass photometry
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In this webinar, Svea Cheeseman, Director of Product Management for Cell and Gene Therapy at Refeyn, discusses how mass photometry can be used to characterize AAV capsids – by measuring the empty/full capsid ratio for AAVs of any serotype with low sample consumption. She also introduces the SamuxMP Auto, Refeyn’s automated mass photometer.
BLOG POST​
Everything you need to know about analysing AAVs with mass photometry
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Read this blog post to learn how mass photometry can streamline your AAV analysis workflows with an easy-to-use benchtop instrument.
More Application Notes
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