ArticleBMC biotechnology2013
High level transient production of recombinant antibodies and antibody fusion proteins in HEK293 cells.
Article in BMC biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 120 papers.
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Who cites it
120 citing papers in PubMed, 197 citations in OpenAlex.
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- Production and Characterization of Recombinant Single-Chain Variable Fragment (scFv) Antibody AgainstInternational journal of molecular sciences · 2026Article
- Enzymatically controlled release of proteins and peptides: A promising, alternative secretion approach.iScience · 2026Article
- The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.Frontiers in bioengineering and biotechnology · 2026Review
- An effective method to enhance transient gene expression by transfecting high-density CHO cells.AMB Express · 2025Article
- A conserved fungal effector disturbs CaNature communications · 2025Article
- Fetal bovine serum: how to leave it behind in the pursuit of more reliable science.Frontiers in toxicology · 2025Review
- A novel PolyAr87-based cell transfection protocol for nanobody expression optimized via a targeted design of transfection approach.Frontiers in bioengineering and biotechnology · 2025Article
- Bispecific antibodies as powerful immunotherapeutic agents for urological cancers: Recent innovations based on preclinical and clinical evidence.International journal of biological sciences · 2025Review
- Unlocking the Puzzle of Mammalian Transfection: The Role of the RNA-sensing-Mediated Interferon Response in the Cellular Defense Against Foreign DNA Intrusion.International journal of biological sciences · 2025Article
- ENPP1/CD203a-targeting heavy-chain antibody reveals cell-specific expression on human immune cells.Cellular and molecular life sciences : CMLS · 2024Article
- Improvement strategies for transient gene expression in mammalian cells.Applied microbiology and biotechnology · 2024Review
- Generation of nanobodies from transgenic 'LamaMice' lacking an endogenous immunoglobulin repertoire.Nature communications · 2024Article
- Studying SARS-CoV-2 interactions using phage-displayed receptor binding domain as a model protein.Scientific reports · 2024Article
- Generation and characterization of antagonistic anti-human CD39 nanobodies.Frontiers in immunology · 2024Article
- Different localization of P2X4 and P2X7 receptors in native mouse lung - lack of evidence for a direct P2X4-P2X7 receptor interaction.Frontiers in immunology · 2024Article
- Induction of long-term tolerance to a specific antigen using anti-CD3 lipid nanoparticles following gene therapy.Molecular therapy. Nucleic acids · 2023Article
- The fatty liver disease-causing protein PNPLA3-I148M alters lipid droplet-Golgi dynamics.bioRxiv : the preprint server for biology · 2023Article
- Molecular reshaping of phage-displayed Interleukin-2 at beta chain receptor interface to obtain potent super-agonists with improved developability profiles.Communications biology · 2023Article
60 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe demand of monospecific high affinity binding reagents, particularly monoclonal antibodies, has been steadily increasing over the last years. Enhanced throughput of antibody generation has been addressed by optimizing in vitro selection using phage display which moved the major bottleneck to the production and purification of recombinant antibodies in an end-user friendly format. Single chain (sc)Fv antibody fragments require additional tags for detection and are not as suitable as immunoglobulins (Ig)G in many immunoassays. In contrast, the bivalent scFv-Fc antibody format shares many properties with IgG and has a very high application compatibility.
resultsIn this study transient expression of scFv-Fc antibodies in human embryonic kidney (HEK) 293 cells was optimized. Production levels of 10-20 mg/L scFv-Fc antibody were achieved in adherent HEK293T cells. Employment of HEK293-6E suspension cells expressing a truncated variant of the Epstein Barr virus (EBV) nuclear antigen (EBNA) 1 in combination with production under serum free conditions increased the volumetric yield up to 10-fold to more than 140 mg/L scFv-Fc antibody. After vector optimization and process optimization the yield of an scFv-Fc antibody and a cytotoxic antibody-RNase fusion protein further increased 3-4-fold to more than 450 mg/L. Finally, an entirely new mammalian expression vector was constructed for single step in frame cloning of scFv genes from antibody phage display libraries. Transient expression of more than 20 different scFv-Fc antibodies resulted in volumetric yields of up to 600 mg/L and 400 mg/L in average.
conclusionTransient production of recombinant scFv-Fc antibodies in HEK293-6E in combination with optimized vectors and fed batch shake flasks cultivation is efficient and robust, and integrates well into a high-throughput recombinant antibody generation pipeline.
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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.