ArticleCytotechnology2002
100-liter transient transfection.
Article in Cytotechnology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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The trial behind it
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
20 citing papers in PubMed.
- Development of novel lipoplex formulation methodologies to improve large-scale transient transfection for lentiviral vector manufacture.Molecular therapy. Methods & clinical development · 2024Article
- Improvement of HEK293 Cell Growth by Adapting Hydrodynamic Stress and Predicting Cell Aggregate Size Distribution.Bioengineering (Basel, Switzerland) · 2023Article
- Recombinant Protein Production and Purification of Insoluble Proteins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- The Emergency Response Capacity of Plant-Based Biopharmaceutical Manufacturing-What It Is and What It Could Be.Frontiers in plant science · 2020Review
- From Farm to Finger Prick-A Perspective on How Plants Can Help in the Fight Against COVID-19.Frontiers in bioengineering and biotechnology · 2020Article
- Article
- The other side of the coin: Leveraging Epstein-Barr virus in research and therapy.Oral oncology · 2016Review
- Optimizing the transient transfection process of HEK-293 suspension cells for protein production by nucleotide ratio monitoring.Cytotechnology · 2014Article
- PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process.Cytotechnology · 2013Article
- Cloning and expression of soluble vascular endothelial growth factors receptor-1 (sFlt-1) fragments in CHO-K1.International journal of clinical and experimental medicine · 2013Article
- Effect of addition of 'carrier' DNA during transient protein expression in suspension CHO culture.Cytotechnology · 2012Article
- Fusion partners can increase the expression of recombinant interleukins via transient transfection in 2936E cells.Protein science : a publication of the Protein Society · 2010Article
- Transient transfection methods for clinical adeno-associated viral vector production.Human gene therapy · 2009Review
- The heterogeneous nature of polyethylenimine-DNA complex formation affects transient gene expression.Cytotechnology · 2009Article
- Transient transfection factors for high-level recombinant protein production in suspension cultured mammalian cells.Molecular biotechnology · 2008Article
- Development of a generic transient transfection process at 100 L scale.Cytotechnology · 2008Article
- Large-scale transfection of mammalian cells for the fast production of recombinant protein.Molecular biotechnology · 2006Review
- Current issues in adeno-associated viral vector production.Gene therapy · 2005Review
- Large-scale transient transfection of mammalian cells: a newly emerging attractive option for recombinant protein production.Journal of structural and functional genomics · 2005Article
- A Study of Recombinant Factor IX in Drosophila Insect S2 Cell Lines Through Transient Gene Expression Technology.Avicenna journal of medical biotechnologyArticle
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This is the first report of two successful 100 l scale transienttransfections in a standard stirred bioreactor. More than half a gram of a monoclonal antibody (IgG) were produced in less than 10 days using a technology called large-scale transient gene expression(LS-TGE). Suspension adapted HEK 293 EBNA SF cells were transfectedwithin a 150 l (nominal) bioreactor by a modified calcium phosphateco-precipitation method with more than 75 mg of plasmid DNA per run.A mixture of three different plasmids, one encoding for the heavychain of a human recombinant immunoglobulin, the other for the corresponding light chain and a third one for the green fluorescent protein (GFP, 2-4% of DNA in transfection cocktail)were co-transfected. The GFP vector was chosen to monitor transfection efficiency. Expression of GFP could be registered asearly as 20 h after DNA addition, using fluorescence microscopy. We demonstrate that transient transfection can be done at the100 l scale, thus providing a new tool to produce hundreds of milligrams or even gram amounts of recombinant protein. Akey advantage of LS-TGE resides in its speed. In the presentedcases, the entire production process for the synthesis of halfa gram of a recombinant antibody, including DNA preparationand necessary expansion of cells prior to transfection, wasexecuted in less than a month. Having an established transfection/expression process allows to run productioncampaigns for any given protein, within one facility, with onesingle host cell line and therefore only one single seed train. Without any need to create and maintain stable cell lines, expression of new r-proteins is not only faster and more economical but also more flexible.
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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.