Abstract
The HEK293 cell line has earned its place as a producer of biotherapeutics. In addition to its ease of growth in serum-free suspension culture and its amenability to transfection, this cell line’s most important attribute is its human origin, which makes it suitable to produce biologics intended for human use. At the present time, the growth and production properties of the HEK293 cell line are inferior to those of non-human cell lines, such as the Chinese hamster ovary (CHO) and the murine myeloma NSO cell lines. However, the modification of genes involved in cellular processes, such as cell proliferation, apoptosis, metabolism, glycosylation, secretion, and protein folding, in addition to bioprocess, media, and vector optimization, have greatly improved the performance of this cell line. This review provides a comprehensive summary of important achievements in HEK293 cell line engineering and on the global engineering approaches and functional genomic tools that have been employed to identify relevant genes for targeted engineering.
Highlights
The human embryonic kidney cell line (HEK293) was created by transforming human embryonic kidney (HEK) cells with sheared fragments of adenovirus type 5 (Ad5) DNA, immortalizing it
Metabolic engineering of HEK293 cells has been concentrated on improving the central carbon metabolism. This has been achieved by restoring the link between glycolysis and the tricarboxylic acid cycle through overexpression of the pyruvate carboxylase (PC) gene [77,78,79,80], and by the knockdown of the pyruvate dehydrogenase kinase gene (PDK), alongside its activator, the hypoxia inducible factor 1 (HIF1) gene [81]
In addition to the production of proteins for therapeutic use, the HEK293 cell line is used for the expression of proteins with proper post-translational modifications (PTMs), such as glycosylation, for structural biology studies [82]
Summary
The human embryonic kidney cell line (HEK293) was created by transforming human embryonic kidney (HEK) cells with sheared fragments of adenovirus type 5 (Ad5) DNA, immortalizing it. Five therapeutic proteins produced in HEK293 cells had been approved by the European Medicines Agency or the U.S Food and Drug Administration for use in humans as of 2015 [13] This cell line has been useful in viral vector production as a result of adenoviral DNA in its genome. It has been adapted to grow in suspension culture in serum-free or chemically defined media and is currently being utilized for recombinant proteins expression in both adherent and suspension platforms. Process and media parameters, such as expression vector, growth media, transfection agent and culture conditions, have been optimized to improved recombinant protein yields from HEK293. This review will discuss the genetic engineering strategies, functional genomics and bioinformatic approaches that have been employed to improve the quality and quantity of recombinant protein production in the HEK293 and its derivative cell lines.
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