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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. ​

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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:

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

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.

To learn how mass photometry can speed up your AAV analytics

How to streamline AAV purification protocols with mass photometry

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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

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.

Image of virus cells superimposed with the title of the webinar "Streamlining AAV characterizsation with automated mass photometry"
Image of a scientist loading sample onto a mass photometer superimposed with the blog title - Everything you need to know about analysing AAVs with mass photometry

BLOG POST​

Everything you need to know about analysing AAVs with mass photometry

Read this blog post to learn how mass photometry can streamline your AAV analysis workflows with an easy-to-use benchtop instrument.

To discuss how mass photometry can improve your AAV analytics workflow

More Application Notes

Browse through our catalogue of application notes highlighting some recent case studies featuring mass photometry.

Investigating lentivirus titer with macro mass photometry

 

Mass photometry supports membrane protein purification protocols 
 

Characterizing protein oligomerization with automated mass photometry

Quantifying heterogeneous AAV capsid loading using mass photometry
 

Download the application note

Download the app note
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