ArticleCytotechnology2008
Development of a generic transient transfection process at 100 L scale.
Article in Cytotechnology, 2008. 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, 50 citations in OpenAlex.
- Development of Lentiviral Packaging Cells and Scale Up of Production to Meet the Growing Demand in Cell and Gene Therapy.Current gene therapy · 2025Review
- Updated transient gene expression protocol in Expi293F cells using PEI.Frontiers in bioengineering and biotechnology · 2025Article
- 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
- Eukaryotic Aggresomes: Protocols and Tips for Their Production, Purification , and Handling.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Lentiviral Vector Bioprocessing.Viruses · 2021Review
- The transient expression of CHIKV VLP in large stirred tank bioreactors.Cytotechnology · 2019Article
- Article
- Enhanced transient recombinant protein production in CHO cells through the co-transfection of the product gene with Bcl-xL.Biotechnology journal · 2014Article
- Effect of Peptone Feeding on Transient Gene Expression Process in CHO DG44.Avicenna journal of medical biotechnology · 2014Article
- Better and faster: improvements and optimization for mammalian recombinant protein production.Current opinion in structural biology · 2014Review
- Optimizing the transient transfection process of HEK-293 suspension cells for protein production by nucleotide ratio monitoring.Cytotechnology · 2014Article
- High level transient production of recombinant antibodies and antibody fusion proteins in HEK293 cells.BMC biotechnology · 2013Article
- Methadone but not morphine inhibits lubiprostone-stimulated Cl- currents in T84 intestinal cells and recombinant human ClC-2, but not CFTR Cl- currents.Cell biochemistry and biophysics · 2013Article
- Expression of recombinant antibodies.Frontiers in immunology · 2013Article
- Environmental parameters influence non-viral transfection of human mesenchymal stem cells for tissue engineering applications.Cell and tissue research · 2012Article
- Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have developed a generic transient transfection process at 100 L scale, using HEK293-EBNA cells and PEI as the transfection reagent for the production of recombinant IgG. The process, including large-scale plasmid preparation, expression at bioreactor scale, capture, purification and, if necessary, endotoxin removal allows reproducible production of more than 0.5 g IgG for in vitro and in vivo studies. We compared the performance of two HEK cell lines, investigated the effect of conditioned medium, optimized the DNA:PEI ratio and implemented a feed strategy to prolong the culture time to increase product yield. The transient transfection protocol developed enables a closed process from seeding culture to protein capture. The challenge of performing a medium exchange before transfection at large scale is solved by applying a continuous centrifugation step between the seeding bioreactor and the production bioreactor. After 7-8 days the harvest and capture is performed in a one-step operation using a Streamline expanded bed chromatography system. Following a polishing step the purified antibody is transferred to the final formulation buffer. The method has shown to be reproducible at 10, 50, and 100 L scale expressing between 5 and 8 mg L(-1) IgG.
Identifiers
What OpenQuestion holds
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.