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Production and characterization of an AAV1-VP3-only capsid: An analytical benchmark standard.

ABSTRACT: Adeno-associated viruses (AAVs) are non-enveloped ssDNA icosahedral T = 1 viruses used as vectors for clinical gene delivery. Currently, there are over 200 AAV-related clinical trials and six approved biologics on the market. As such new analytical methods are continually being developed to characterize and monitor the quality and purity of manufactured AAV vectors, these include ion-exchange chromatography and Direct Mass Technology. However, these methods require homogeneous analytical standards with a high molecular weight standard comparable to the mass of an AAV capsid. Described here is the design, production, purification, characterization, and the cryo-electron microscopy structure of an AAV1-VP3-only capsid that fulfills this need as a calibrant to determine capsid mass, charge, homogeneity, and transgene packaging characteristics.

SUBMITTER: Mietzsch M

PROVIDER: S-EPMC10238842 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature

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Production and characterization of an AAV1-VP3-only capsid: An analytical benchmark standard.

Mietzsch Mario M Liu Weijing W Ma Ke K Bennett Antonette A Nelson Austin R AR Gliwa Keely K Chipman Paul P Fu Xiaofeng X Bechler Shane S McKenna Robert R Viner Rosa R

Molecular therapy. Methods & clinical development 20230509

Adeno-associated viruses (AAVs) are non-enveloped ssDNA icosahedral T = 1 viruses used as vectors for clinical gene delivery. Currently, there are over 200 AAV-related clinical trials and six approved biologics on the market. As such new analytical methods are continually being developed to characterize and monitor the quality and purity of manufactured AAV vectors, these include ion-exchange chromatography and Direct Mass Technology. However, these methods require homogeneous analytical standar ...[more]

PMID: 37273903

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