ArticleBiotechnology and bioengineering2022
Only a small fraction of cells produce assembled capsids during transfection-based manufacturing of adeno-associated virus vectors.
Article in Biotechnology and bioengineering, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- AAV vector production in suspension cells using PEI transfection and sodium butyrate with orthogonal assessment of function and quality.Molecular therapy. Advances · 2026Article
- Comparative Transcriptomic Profiling Reveals Differences in Initiation of Antiviral Response in Low rAAV Producing HEK293 Suspension Cells.Biotechnology journal · 2026Article
- Comparative Analysis of rAAV Production from Plasmid-Encoded Versus Chromosomally Integrated rAAV Transgene in HEK293 Cells.International journal of molecular sciences · 2026Article
- Trivalent ions kinetic-gating for producing high-concentration and shelf-stable plasmid DNA/PEI particles.Nature communications · 2026Article
- AAV Assembled Capsids Are Produced in Cells Blocked From Cell Cycle Progression.Biotechnology and bioengineering · 2026Article
- Heterogeneity in an adeno-associated virus transfection-based production process limits the production efficiency.Scientific reports · 2025Article
- Accelerated Adeno Associated Virus Upstream Process Development From High-Throughput Systems to Clinical Scale.Biotechnology and bioengineering · 2025Article
- Buffer Valency Engineering Enables High-concentration and Shelf-stable DNA Transfection Particles for Viral Vector Production.Research square · 2025Article
- AAV-Based Gene Therapy: Opportunities, Risks, and Scale-Up Strategies.International journal of molecular sciences · 2025Review
- AAVone: A cost-effective, single-plasmid solution for efficient AAV production with reduced DNA impurities.Molecular therapy. Nucleic acids · 2025Article
- Model-Based Characterization of the Metabolism of Recombinant Adeno-Associated Virus (rAAV) Production via Human Embryonic Kidney (HEK293) Cells.Bioengineering (Basel, Switzerland) · 2025Article
- PCR-based analytics of gene therapies using adeno-associated virus vectors: Considerations for cGMP method development.Molecular therapy. Methods & clinical development · 2023Review
- Design space determination to optimize DNA complexation and full capsid formation in transient rAAV manufacturing.Biotechnology and bioengineering · 2023Article
- Comparison of three AUC techniques for the determination of the loading status and capsid titer of AAVs.European biophysics journal : EBJ · 2023Article
- AAV genome modification for efficient AAV production.Heliyon · 2023Review
- Only a small fraction of cells produce assembled capsids during transfection-based manufacturing of adeno-associated virus vectors.Biotechnology and bioengineering · 2022Article
- Status and future of recombinant adeno-associated virus vector manufacturing.Biotechnology progressReview
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Plasmid transfection of mammalian cells is the dominant platform used to produce adeno-associated virus (AAV) vectors for clinical and research applications. Low yields from this platform currently make it difficult to supply these activities with adequate material. In an effort to better understand the current limitations of transfection-based manufacturing, this study examines what proportion of cells in a model transfection produce appreciable amounts of assembled AAV capsid. Using conformation-specific antibody staining and flow cytometry, we report the surprising result that despite obtaining high transfection efficiencies and nominal vector yields in our model system, only 5%-10% of cells appear to produce measurable levels of assembled AAV capsids. This finding implies that considerable increases in vector titer could be realized through increasing the proportion of productive cells. Furthermore, we suggest that the flow cytometry assay used here to quantify productive cells may be a useful metric for future optimization of transfection-based AAV vector manufacturing platforms.
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