ArticleJournal of virology2006
Scalable generation of high-titer recombinant adeno-associated virus type 5 in insect cells.
Article in Journal of virology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 58 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
58 citing papers in PubMed.
- Variation of VP2 stoichiometry and deamidation of VP1 during production and their impacts on the transduction efficiency of AAV vectors.Molecular therapy. Methods & clinical development · 2025Article
- Intensification of rAAV Production Based on HEK293 Cell Transient Transfection.Biotechnology journal · 2025Article
- Impact of multiplicity of infection and baculovirus co-infection ratio on recombinant adeno-associated virus vector production in insect cells.Frontiers in bioengineering and biotechnology · 2025Article
- Production of recombinant adeno-associated virus 5 using a novel self-attenuating adenovirus production platform.Molecular therapy. Methods & clinical development · 2024Article
- A robust and flexible baculovirus-insect cell system for AAV vector production with improved yield, capsid ratios and potency.Molecular therapy. Methods & clinical development · 2024Article
- A Direct Comparison of rAAV5 Variants Derived from the Baculovirus Expression System Using LC-MS Workflows Demonstrates Key Differences in Overall Production Yield, Product Quality and Vector Efficiency.International journal of molecular sciences · 2024Article
- Advanced biomanufacturing and evaluation of adeno-associated virus.Journal of biological engineering · 2024Article
- Engineered CHO cells as a novel AAV production platform for gene therapy delivery.Scientific reports · 2023Article
- Stowaways in the cargo: Contaminating nucleic acids in rAAV preparations for gene therapy.Molecular therapy : the journal of the American Society of Gene Therapy · 2023Review
- Mechanistic modeling explains the production dynamics of recombinant adeno-associated virus with the baculovirus expression vector system.Molecular therapy. Methods & clinical development · 2023Article
- Synthetic Biology Design as a Paradigm Shift toward Manufacturing Affordable Adeno-Associated Virus Gene Therapies.ACS synthetic biology · 2023Review
- Development of an insect cell-based adeno-associated virus packaging cell line employing advancedMolecular therapy. Methods & clinical development · 2022Article
- Utility of Alternative Promoters for Foreign Gene Expression Using the Baculovirus Expression Vector System.Viruses · 2022Article
- Article
- Process improvement of adeno-associated virus (AAV) production.Frontiers in chemical engineering · 2022Article
- Detailed Protocol for the Novel and Scalable Viral Vector Upstream Process for AAV Gene Therapy Manufacturing.Human gene therapy · 2021Article
- Anion-exchange HPLC assay for separation and quantification of empty and full capsids in multiple adeno-associated virus serotypes.Molecular therapy. Methods & clinical development · 2021Article
- Adeno-associated virus capsid assembly is divergent and stochastic.Nature communications · 2021Article
- Contribution to HIV Prevention and Treatment by Antibody-Mediated Effector Function and Advances in Broadly Neutralizing Antibody Delivery by Vectored Immunoprophylaxis.Frontiers in immunology · 2021Review
- New Directions in Pulmonary Gene Therapy.Human gene therapy · 2020Review
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
We established a method for production of recombinant adeno-associated virus type 5 (rAAV5) in insect cells by use of baculovirus expression vectors. One baculovirus harbors a transgene between the inverted terminal repeat sequences of type 5, and the second expresses Rep78 and Rep52. Interestingly, the replacement of type 5 Rep52 with type 1 Rep52 generated four times more rAAV5 particles. We replaced the N-terminal portion of type 5 VP1 with the equivalent portion of type 2 to generate infectious AAV5 particles. The rAAV5 with the modified VP1 required alpha2-3 sialic acid for transduction, as revealed by a competition experiment with an analog of alpha2-3 sialic acid. rAAV5-GFP/Neo with a 4.4-kb vector genome produced in HEK293 cells or Sf9 cells transduced COS cells with similar efficiencies. Surprisingly, Sf9-produced humanized Renilla green fluorescent protein (hGFP) vector with a 2.4-kb vector genome induced stronger GFP expression than the 293-produced one. Transduction of murine skeletal muscles with Sf9-generated rAAV5 with a 3.4-kb vector genome carrying a human secreted alkaline phosphatase (SEAP) expression cassette induced levels of SEAP more than 30 times higher than those for 293-produced vector 1 week after injection. Analysis of virion DNA revealed that in addition to a 2.4- or 3.4-kb single-stranded vector genome, Sf9-rAAV5 had more-abundant forms of approximately 4.7 kb, which appeared to correspond to the monomer duplex form of hGFP vector or truncated monomer duplex SEAP vector DNA. These results indicated that rAAV5 can be generated in insect cells, although the difference in incorporated virion DNA may induce different expression patterns of the transgene.
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Registered trials
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