ArticleBiotechnology and bioengineering2008
Serum-free transient protein production system based on adenoviral vector and PER.C6 technology: high yield and preserved bioactivity.
Article in Biotechnology and bioengineering, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
18 citing papers in PubMed, 43 citations in OpenAlex.
- Production and Storage of High-Capacity Adenoviral Vectors for the Delivery of Advanced CRISPR Systems.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Precision genome editing using combinatorial viral vector delivery of CRISPR-Cas9 nucleases and donor DNA constructs.Nucleic acids research · 2025Article
- Adenovirus vector-based vaccines as forefront approaches in fighting the battle against flaviviruses.Human vaccines & immunotherapeutics · 2022Review
- Broadening the reach and investigating the potential of prime editors through fully viral gene-deleted adenoviral vector delivery.Nucleic acids research · 2021Article
- Affecting HEK293 Cell Growth and Production Performance by Modifying the Expression of Specific Genes.Cells · 2021Review
- Applications and analysis of hydrolysates in animal cell culture.Bioresources and bioprocessing · 2021Review
- Integrating gene delivery and gene-editing technologies by adenoviral vector transfer of optimized CRISPR-Cas9 components.Gene therapy · 2020Article
- Review
- Article
- Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives.Critical reviews in biotechnology · 2016Review
- Adenoviral vectors encoding CRISPR/Cas9 multiplexes rescue dystrophin synthesis in unselected populations of DMD muscle cells.Scientific reports · 2016Article
- Adenoviral vector-based strategies against infectious disease and cancer.Human vaccines & immunotherapeutics · 2016Review
- Selection-free gene repair after adenoviral vector transduction of designer nucleases: rescue of dystrophin synthesis in DMD muscle cell populations.Nucleic acids research · 2016Article
- Adenoviral vector DNA for accurate genome editing with engineered nucleases.Nature methods · 2014Article
- Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells.Scientific reports · 2014Article
- Polyethyleneimine-based transient gene expression processes for suspension-adapted HEK-293E and CHO-DG44 cells.Protein expression and purification · 2013Article
- Differential integrity of TALE nuclease genes following adenoviral and lentiviral vector gene transfer into human cells.Nucleic acids research · 2013Article
- Recent advances in technology supporting biopharmaceutical production from mammalian cells.Applied microbiology and biotechnology · 2012Review
Corrections and comments
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Authors and funding
13 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Stable E1 transformed cells, like PER.C6, are able to grow at scale and to high cell densities. E1-deleted adenoviruses replicate to high titer in PER.C6 cells whereas subsequent deletion of E2A from the vector results in absence of replication in PER.C6 cells and drastically lowers the expression of adenovirus proteins in such cells. We therefore considered the use of an DeltaE1/DeltaE2 type 5 vector (Ad5) to deliver genes to PER.C6 cells growing in suspension with the aim to achieve high protein yield. To evaluate the utility of this system we constructed DeltaE1/DeltaE2 vector carrying different classes of protein, that is, the gene coding for spike protein derived from the Coronavirus causing severe acute respiratory syndrome (SARS-CoV), a gene coding for the SARS-CoV receptor or the genes coding for an antibody shown to bind and neutralize SARS-CoV (SARS-AB). The DeltaE1/DeltaE2A-vector backbones were rescued on a PER.C6 cell line engineered to constitutively over express the Ad5 E2A protein. Exposure of PER.C6 cells to low amounts (30 vp/cell) of DeltaE1/DeltaE2 vectors resulted in highly efficient (>80%) transduction of PER.C6 cells growing in suspension. The efficient cell transduction resulted in high protein yield (up to 60 picogram/cell/day) in a 4 day batch production protocol. FACS and ELISA assays demonstrated the biological activity of the transiently produced proteins. We therefore conclude that DeltaE1/DeltaE2 vectors in combination with the PER.C6 technology may provide a viable answer to the increasing demand for high quality, high yield recombinant protein.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.