ArticleMolecular biotechnology2008
Transient transfection factors for high-level recombinant protein production in suspension cultured mammalian cells.
Article in Molecular biotechnology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 95 citations in OpenAlex.
- High-efficiency transfection of Acanthamoeba castellanii using a cationic polymer.Cell reports methods · 2026Article
- An effective method to enhance transient gene expression by transfecting high-density CHO cells.AMB Express · 2025Article
- Accelerated Adeno Associated Virus Upstream Process Development From High-Throughput Systems to Clinical Scale.Biotechnology and bioengineering · 2025Article
- Advances in Serum-Free Suspension Culture Technology for Animal Cells and Their Applications.Vaccines · 2025Review
- A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology.Antibodies (Basel, Switzerland) · 2025Article
- A transposon-based transient transfection system in CHO-K1 cells enables quality prediction of stable cell line proteins.Bioprocess and biosystems engineering · 2025Article
- MiRNA Chaining for Efficient Stable Overexpression to Improve Protein Quantity and Quality in CHO Cells.Methods in molecular biology (Clifton, N.J.) · 2025Article
- A novel PolyAr87-based cell transfection protocol for nanobody expression optimized via a targeted design of transfection approach.Frontiers in bioengineering and biotechnology · 2025Article
- Review
- Improvement strategies for transient gene expression in mammalian cells.Applied microbiology and biotechnology · 2024Review
- Promoting the production of challenging proteins via induced expression in CHO cells and modified cell-free lysates harboring T7 RNA polymerase and mutant eIF2α.Synthetic and systems biotechnology · 2024Article
- Stable overexpression of native and artificial miRNAs for the production of differentially fucosylated antibodies in CHO cells.Engineering in life sciences · 2024Article
- Article
- Long-Term Culturing of FreeStyle 293-F Cells Affects Immunoglobulin G Glycome Composition.Biomolecules · 2023Article
- Exploring Parametric and Mechanistic Differences between Expi293FAntibodies (Basel, Switzerland) · 2023Article
- Augmenting recombinant antibody production in HEK293E cells: optimizing transfection and culture parameters.Antibody therapeutics · 2022Article
- Novel human cell expression method reveals the role and prevalence of posttranslational modification in nonmuscle tropomyosins.The Journal of biological chemistry · 2021Article
- Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications.Diagnostics (Basel, Switzerland) · 2021Article
- Solubility, Stability, and Avidity of Recombinant Antibody Fragments Expressed in Microorganisms.Frontiers in microbiology · 2020Review
- A simple, rapid method for evaluation of transfection efficiency based on fluorescent dye.Bioengineered · 2017Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The efficient transfection of cloned genes into mammalian cells system plays a critical role in the production of large quantities of recombinant proteins (r-proteins). In order to establish a simple and scaleable transient protein production system, we have used a cationic lipid-based transfection reagent-FreeStyle MAX to study transient transfection in serum-free suspension human embryonic kidney (HEK) 293 and Chinese hamster ovary (CHO) cells. We used quantification of green fluorescent protein (GFP) to monitor transfection efficiency and expression of a cloned human IgG antibody to monitor r-protein production. Parameters including transfection reagent concentration, DNA concentration, the time of complex formation, and the cell density at the time of transfection were analyzed and optimized. About 70% GFP-positive cells and 50-80 mg/l of secreted IgG antibody were obtained in both HEK-293 and CHO cells under optimal conditions. Scale-up of the transfection system to 1 l resulted in similar transfection efficiency and protein production. In addition, we evaluated production of therapeutic proteins such as human erythropoietin and human blood coagulation factor IX in both HEK-293 and CHO cells. Our results showed that the higher quantity of protein production was obtained by using optimal transient transfection conditions in serum-free adapted suspension mammalian cells.
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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.