Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes
Open accessAbstract
The generation of clinical good manufacturing practices (GMP)-grade adeno-associated virus (AAV) vectors requires purification strategies that support the generation of vectors of high purity, and that exhibit a good safety and efficacy profile. To date, most reported purification schemas are serotype dependent, requiring method development for each AAV gene therapy product. Here, we describe a platform purification process that is compatible with the purification of multiple AAV serotypes. The method generates vector preparations of high purity that are enriched for capsids with full vector genomes, and that minimizes the fractional content of empty capsids. The two-column purification method, a combination of affinity and ion exchange chromatographies, is compatible with a range of AAV serotypes generated by either the transient triple transfection method or the more scalable producer cell line platform. In summary, the adaptable purification method described can be used for the production of a variety of high-quality AAV vectors suitable for preclinical testing in animal models of diseases.
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AUC is limited to purified samples without residual impurities or excipients and requires prior titration into the linear range 2 × 10^12-5 × 10^12 cp/mL.
“Sensitivity of the detectors and the risk of induced density gradients limit AUC to purified samples without residual impurities or excipients, and prior titration into the small linear range of the detector, 2 × 1012–5 × 1012 cp/mL, is needed.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [51]
AUC requires large sample volumes (400-500 μL) at high concentrations around 2 × 10^12-5 × 10^12 vg/mL, representing significant loss of unrecoverable material during early production stages.
“In general, AUC requires large sample volumes, 400–500 μL, at high concentrations around 2 × 1012–5 × 1012 vg/mL, representing a significant loss of unrecoverable material during early production stages.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [51]
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- Authors
- Nass, S. A., Mattingly, M. A., Woodcock, D. A., Burnham, B. L., Ardinger, J. A., Osmond, S. E., Frederick, A. M., Scaria, A., Cheng, S. H., & O’Riordan, C. R.
- Corresponding author
- O’Riordan, Catherine R.
- Venue
- Molecular Therapy - Methods & Clinical Development
- Publisher
- Elsevier BV
- Published
- 2018-06
- Volume
- 9
- Pages
- 33-46
- ISSN
- 2329-0501 (ISSN-L)
- PMC
- PMC5767896
- OpenAlex
- W2781108704
- Cited by
- 146
- References
- 27
- Citations by year
- 2026: 8 · 2025: 24 · 2024: 16 · 2023: 29 · 2022: 13 · 2021: 27 · 2020: 17 · 2019: 6 · 2018: 6
- FWCI
- 5.26
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecular Therapy — Methods & Clinical Development (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- Sanofi
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- adeno-associated virus vectors, AAV serotypes, vector purification, affinity chromatography, ion-exchange chromatography, producer cell lines
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