Improving the Quality of Adeno-Associated Viral Vector Preparations: The Challenge of Product-Related Impurities
Abstract
Adeno-associated viral (AAV) vectors have emerged as one of the most popular gene transfer systems in both research and clinical gene therapy. As AAV vectors are derived from a stealth, nonpathogenic virus and lack active integrase activity, these vectors are frequently applied for in vivo gene therapy of liver, muscle, and other postmitotic tissues. Although long-term transgene expression from AAV vector episomes is reported from these tissues, the episomal nature of AAV-once regarded as disadvantage-has become an attractive feature for gene-editing approaches targeting proliferating cells. In response to the high demand, AAV vector production is receiving special attention. Besides particle yields and biological activity, the most important concern is improving vector purity. The most difficult task in this regard is removal of defective particles, that is, capsids that are either empty or contain DNA other than the full-length vector genomes. Herein, we characterize and discuss these so-called product-related impurities, methods for their detection, as well as strategies to avoid or reduce their formation.
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Empty capsids, as well as capsids with non-transgene sequences, can elicit an unnecessary immune response and compete for vector binding sites, increasing the risk of immunotoxicity and lowering the rate of transduction of full capsids required for product efficacy.
“Empty capsids, as well as capsids with non-transgene sequences, can elicit an unnecessary immune response and compete for vector binding sites, increasing the risk of immunotoxicity and lowering the rate of transduction of full capsids, which are required for product efficacy.28,30”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [30]
Other impurities besides empty and partially filled capsids, such as residual helper viruses, residual proteins, or nucleic acids, can contribute to immuno- or genotoxic effects.
“Other impurities, besides empty and partially filled capsids, can contribute to immuno- or genotoxic effects, e.g., residual helper viruses (e.g., from upstream transduction steps), residual proteins (e.g., from the host cell or the cell culture medium), or nucleic acids (e.g., host cell DNA/RNA).”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [30]
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- Authors
- Schnödt, M., & Büning, H.
- Affiliations
- Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany.
- University of Cologne (DE)
- ORCID
- Büning, Hildegard
- Venue
- Human Gene Therapy Methods
- Publisher
- SAGE Publications
- Published
- 2017-06
- Volume
- 28
- Issue
- 3
- Pages
- 101-108
- ISSN
- 1946-6536 (ISSN-L)
- 1946-6544
- OpenAlex
- W2607401554
- Cited by
- 77
- References
- 81
- Citations by year
- 2026: 5 · 2025: 13 · 2024: 10 · 2023: 18 · 2022: 5 · 2021: 7 · 2020: 7 · 2019: 10 · 2018: 2
- FWCI
- 0.67
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Human Gene Therapy Methods (publishedVersion)
- PubMed (publishedVersion)
- Kölner Universitäts PublikationsServer (Universität zu Köln) (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- adeno-associated virus vectors, AAV vector production, product-related impurities, empty capsids, gene therapy vectors, genome editing
- MeSH terms
- Dependovirus, Animals, Genetic Vectors, Humans, Genetic Therapy, DNA Contamination
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