Affecting HEK293 Cell Growth and Production Performance by Modifying the Expression of Specific Genes

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.

HEK293 Cell Line as a Platform to Produce Recombinant Proteins and Viral Vectors

“…Similarly, the HEK293-6E cell line was established by the expression of a truncated EBNA-1, lacking Gly-Gly-Ala domain. HEK293-6E showed improved transient gene expression and cell growth compared to HEK293-E (Abaandou et al, 2021).…”

“…The use of the HEK293 host cell eliminates issues of potential immunogenicity due to the presence of non-human PTMs ( Durocher and Butler, 2009 ). Its ease of transfectability and relatively high protein productivity makes it a popular choice for small scale production of recombinant proteins for scientific studies ( Abaandou et al, 2021 ) and its capability to adapt to a suspension free culture facilitates its use in large scale biotherapeutics production ( Baldi et al, 2005 ; Dumont et al, 2015 ).…”

Expressed therapeutic protein yields are predicted by transiently transfected mammalian cell population

“…Although the amount of total expressed protein (as shown in Figure2) may be intuitively correct way to estimate transfection performance, long-term protein production is also dependent on another important aspect-successful division of transfected cells. Each transfection-positive cell clone gives rise to approximately 100 to 500 protein-producing offspring, depending on the cell line and cultivation conditions [13]. For example, a cell that is successfully transfected (estimated at early time point, such as in Figure2) but is unable to undergo cell division and produce daughter cells (which will only be apparent after several days post-transfection, such as in Figure1b) is ultimately not contributing to expressed protein yields.…”

Novel cell lines derived from Chinese hamster kidney tissue

“…In the long history of tissue-derived cell line establishment, many cell lines are of kidney origin [ 1 ]. Kidney-derived cell lines have been widely used in biotechnology fields such as virus production [ 2 ], vaccine production [ 2 ], recombinant protein production [ 2 , 3 ], and drug screening using their membrane permeability properties [ 4 ], and have also been applied in in vitro studies of physiological processes such as osmoregulation and excretion [ 5 ]. In the development of renal cell lines such as 293 cells [ 6 ], BHK cells [ 7 ], and CV-1 cells [ 8 ], transformation and immortalization were achieved by viral infection and the introduction of genes encoding viral constituent proteins.…”