Recombinant proteins expressed in HEK293 cells. DOI:10.1093/dnares/dsi010
HEK293 cells (Human Embryonic Kidney 293 cells are among the most widely used cell lines in biotechnology and biomedical research. First isolated in the 1970s, this cell line was derived by transfecting human embryonic kidney cells with adenovirus type 5 DNA.
History and Development of HEK293 Cells
The "293" in the cell line’s name references the 293rd experiment conducted in the series that successfully yielded the immortalized cell line. These cells gained immortalization through the incorporation of adenoviral genes, allowing continuous growth in laboratory conditions.
While originally thought to derive solely from kidney cells, more recent studies suggest that the adenovirus preferentially transformed cells of neuronal lineage present in the original kidney tissue culture.
Human Origin: The human derivation of HEK293 cells ensures that the expressed proteins have human-like post-translational modifications (PTMs), such as glycosylation, essential for therapeutic proteins and biologics intended for human use.
High Transfection Efficiency: HEK293 cells are highly amenable to transfection methods, including calcium phosphate and polyethyleneimine (PEI), enabling the efficient introduction of foreign DNA for gene expression studies and recombinant protein production.
Growth Versatility: These cells grow well in both adherent and suspension cultures. When adapted to serum-free, chemically defined media, HEK293 cells can achieve high cell densities, making them suitable for large-scale production.
Applications of HEK293 Cells in Biopharmaceuticals
Recombinant Protein Production
HEK293 cells are widely used to produce therapeutic proteins that require human-like PTMs for proper function. Examples include clotting factors, natural anticoagulants, and complex antibody fragments. For example, therapies such as Eloctate (a recombinant Factor VIII Fc fusion protein) and Trulicity (a GLP-1 receptor agonist) are produced using HEK293 derivatives.
Gene Therapy and Viral Vector Production
HEK293 cells are the leading platform for producing viral vectors, such as adeno-associated viruses (AAVs) and lentiviral vectors. Their adaptability to high-density suspension culture and ease of transfection make them ideal for high-titer production.
Research Applications
HEK293 cells are a go-to model for studying:
Protein-protein interactions
Post-translational modifications of proteins
Ion channel activity and drug screening
Genetic editing using CRISPR and other tools
HEK293 vs. CHO Cells
Both HEK293 and CHO cells are mammalian cell platforms widely used in the biotech industry, but their applications differ.
CHO cells are known for scalability, robustness, and cost-effectiveness, CHO cells dominate the production of monoclonal antibodies and other high-yield biologics. HEK293 cells are preferred for biologics requiring human PTMs or viral vector production. HEK293 cells are more versatile for challenging protein targets.
Improved Glycosylation: Knockouts of glycosylation-related genes and the use of glycoengineering tools are enabling more consistent glycan profiles.
Stable Producer Lines: Efforts to create stable HEK293-based lines for viral vector production are reducing reliance on transient transfection methods.
Process Optimization: Advances in media formulations and bioreactor technology are improving cell growth and protein yields.
Expression systems utilizing mammalian cells for recombinant protein manufacture introduces proper protein folding, post-translational modifications, and protein assembly, which are important for producing proteins of complete biological activity, and thus is in high demand in the therapeutic development and application fields, especially for biotechnology and pharmaceutical companies. Besides industry use, academic research groups can also rely on recombinant mammalian proteins for functional studies and high-resolution structure determination.
Biointron, with high flexibility, good cost-effectiveness, and large production capabilities, offers high-quality protein expression service via mammalian cell system using CHO-K1 and HEK293 cells.
References:
Tan, E., Hui Chin, C. S., Sherman Lim, Z. F., & Ng, S. K. (2021). HEK293 Cell Line as a Platform to Produce Recombinant Proteins and Viral Vectors. Frontiers in Bioengineering and Biotechnology, 9, 796991. https://doi.org/10.3389/fbioe.2021.796991
Abaandou, L., Quan, D., & Shiloach, J. (2021). Affecting HEK293 Cell Growth and Production Performance by Modifying the Expression of Specific Genes. Cells, 10(7), 1667. https://doi.org/10.3390/cells10071667
Dumont, J., Euwart, D., Mei, B., Estes, S., & Kshirsagar, R. (2015). Human cell lines for biopharmaceutical manufacturing: History, status, and future perspectives. Critical Reviews in Biotechnology, 36(6), 1110. https://doi.org/10.3109/07388551.2015.1084266
