Expression of UCOE and HSP27 molecular elements to improve the stable
protein production on HEK293 cells.
Abstract
Recombinant proteins represent one of the greatest achievements of
modern pharmaceutical biotechnology, as they are increasingly used in
almost all branches of medicine to treat a wide range of conditions. For
responding to this demand, different cell engineering approaches have
been developed to improve their expression. Some of them include the use
of genetic elements that have the ability to prevent the silencing of
the gene of interest and the generation of resistant cell lines to
inhibit or avoid the Programmed Cell Death (PCD). This research focuses
on the analysis of the effect of overexpression of UCOE elements and the
HSP27 protein individually and together on the production of human rIFNγ
in HEK293 cells. Our results show that UCOE elements of 4 kbp do not
have any effect on protein production in HEK293 cells while
overexpression of HSP27 prolongs the stationary phase during growth
kinetics. It was also observed that the Qp of rIFNγ is 96-fold higher in
clones containing the HSP27/UCOE combination compared to the clone
containing only UCOE elements nor to the control cell HEK293. These
results correlate with the MCP analyses, as HSP27 overexpression was
observed to decrease the expression of bax, caspase 3 and cytochrome C
messengers, beclin and LC3II. This study proposes the utility of a cell
engineering approach based on the overexpression of the human HSP27
protein to develop the production of recombinant viruses and proteins in
HEK293 cells.