<div class="csl-bib-body">
<div class="csl-entry">Wagner, C., Fuchsberger, F. F., Innthaler, B., Lemmerer, M., & Birner-Grünberger, R. (2023). Quantification of Empty, Partially Filled and Full Adeno-Associated Virus Vectors Using Mass Photometry. <i>International Journal of Molecular Sciences</i>, <i>24</i>(13), Article 11033. https://doi.org/10.3390/ijms241311033</div>
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dc.identifier.issn
1661-6596
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/188041
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dc.description.abstract
Adeno-associated viruses (AAV) are one of the most commonly used vehicles in gene therapies for the treatment of rare diseases. During the AAV manufacturing process, particles with little or no genetic material are co-produced alongside the desired AAV capsid containing the transgene of interest. Because of the potential adverse health effects of these byproducts, they are considered impurities and need to be monitored carefully. To date, analytical ultracentrifugation (AUC), transmission electron microscopy (TEM) and charge-detection mass spectrometry (CDMS) are used to quantify these subspecies. However, they are associated with long turnaround times, low sample throughput and complex data analysis. Mass photometry (MP) is a fast and label-free orthogonal technique which is applicable to multiple serotypes without the adaption of method parameters. Furthermore, it can be operated with capsid titers as low as 8 × 1010 cp mL-1 with a CV < 5% using just 10 µL total sample volume. Here we demonstrate that mass photometry can be used as an orthogonal method to AUC to accurately quantify the proportions of empty, partially filled, full and overfull particles in AAV samples, especially in cases where ion-exchange chromatography yields no separation of the populations. In addition, it can be used to confirm the molar mass of the packaged genomic material in filled AAV particles.
en
dc.language.iso
en
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dc.publisher
MDPI
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dc.relation.ispartof
International Journal of Molecular Sciences
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dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
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dc.subject
Capsid
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dc.subject
Capsid Proteins
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dc.subject
Microscopy, Electron, Transmission
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dc.subject
adeno-associated virus vectors
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dc.subject
analytical ultracentrifugation
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dc.subject
genomic cargo
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dc.subject
partially filled particles
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dc.subject
single-molecule mass photometry
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dc.subject
Dependovirus
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dc.subject
Genetic Vectors
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dc.title
Quantification of Empty, Partially Filled and Full Adeno-Associated Virus Vectors Using Mass Photometry