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Quantification of Empty, Partially Filled and Full Adeno-Associated Virus Vectors Using Mass Photometry.
Wagner, Christina; Fuchsberger, Felix F; Innthaler, Bernd; Lemmerer, Martin; Birner-Gruenberger, Ruth.
Affiliation
  • Wagner C; Analytical Development Europe, Takeda Vienna, 1220 Vienna, Austria.
  • Fuchsberger FF; Gene Therapy Process Development Europe, Takeda Orth an der Donau, 2304 Orth an der Donau, Austria.
  • Innthaler B; Gene Therapy Process Development Europe, Takeda Orth an der Donau, 2304 Orth an der Donau, Austria.
  • Lemmerer M; Analytical Development Europe, Takeda Vienna, 1220 Vienna, Austria.
  • Birner-Gruenberger R; Institute of Chemical Technologies and Analytics, Technische Universität Wien, 1040 Vienna, Austria.
Int J Mol Sci ; 24(13)2023 Jul 03.
Article in En | MEDLINE | ID: mdl-37446211
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Dependovirus / Genetic Vectors Type of study: Risk_factors_studies Language: En Journal: Int J Mol Sci Year: 2023 Document type: Article Affiliation country: Austria

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Dependovirus / Genetic Vectors Type of study: Risk_factors_studies Language: En Journal: Int J Mol Sci Year: 2023 Document type: Article Affiliation country: Austria