ArticlePloS one2015
Enhancing Protein Expression in HEK-293 Cells by Lowering Culture Temperature.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed.
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- Low-temperature enhances production of severe fever with thrombocytopenia syndrome virus virus-like particles.Applied microbiology and biotechnology · 2025Article
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- Structural Insights into Partial Activation of the Prototypic G Protein-Coupled Adenosine AACS pharmacology & translational science · 2024Article
- A solution for highly efficient electroporation of primary cytotoxic T lymphocytes.BMC biotechnology · 2024Article
- Investigating dynamics of lentiviral vector secretion from HEK293T producer cells using a fractionated perfusion system.Biotechnology journal · 2024Article
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- Adaptive Laboratory Evolution to Improve Recombinant Protein Production Using Insect Cells.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Multiparametric domain insertional profiling of adeno-associated virus VP1.Molecular therapy. Methods & clinical development · 2023Article
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- Co-Surfactant-Free Bioactive Protein Nanosheets for the Stabilization of Bioemulsions Enabling Adherent Cell Expansion.Biomacromolecules · 2023Article
- Cell-cycle arrest at the G1/S boundary enhances transient voltage-gated ion channel expression in human and insect cells.Cell reports methods · 2023Article
- Improving expression and assembly of difficult-to-express heterologous proteins in Saccharomyces cerevisiae by culturing at a sub-physiological temperature.Microbial cell factories · 2023Article
- Comparison of vector elements and process conditions in transient and stable suspension HEK293 platforms using SARS-CoV-2 receptor binding domain as a model protein.BMC biotechnology · 2023Article
- Flow Cytometry-Based Detection of Siglec Ligands.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Enhancing productivity of Chinese hamster ovary (CHO) cells: synergistic strategies combining low-temperature culture and mTORC1 signaling engineering.Frontiers in bioengineering and biotechnology · 2023Article
- Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing.Viruses · 2022Article
- Augmenting recombinant antibody production in HEK293E cells: optimizing transfection and culture parameters.Antibody therapeutics · 2022Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Animal cells and cell lines, such as HEK-293 cells, are commonly cultured at 37°C. These cells are often used to express recombinant proteins. Having a higher expression level or a higher protein yield is generally desirable. As we demonstrate in this study, dropping culture temperature to 33°C, but not lower, 24 hours after transient transfection in HEK-293S cells will give rise to ~1.5-fold higher expression of green fluorescent protein (GFP) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. By following the time course of the GFP-expressing cells growing at 37°C and 33°C from 24 hours after transfection (including 19 hours recovery at 37°C in the normal growth medium), we found that a mild hypothermia (i.e., 33°C) reduces the growth rate of HEK-293S cells, while increasing cellular productivity of recombinant proteins. As a result, green cells remain undivided in a longer period of time. Not surprisingly, the property of a recombinant protein expressed in the cells grown at 33°C is unaffected, as shown by the use of AMPA receptors. We further demonstrate with the use of PC12 cells that this method may be especially useful when a recombinant protein is difficult to express using a chemical-based, transient transfection method.
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