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 across nearly all branches of medicine to treat a wide range of conditions. In response to this demand, various cell engineering approaches have been developed to improve their expression. Some of these approaches involve the use of genetic elements that prevent the silencing of the gene of interest, as well as the generation of resistant cell lines to inhibit or avoid programmed cell death (PCD). This research focuses on analyzing the effects of overexpression of UCOE elements and the HSP27 protein, both individually and together, on the production of human rIFNγ in HEK293 cells. Our results show that 4-Kb UCOE elements have no effect on protein production in HEK293 cells, while overexpression of HSP27 prolongs the stationary phase during growth kinetics. The Qp of rIFNγ is 96-fold higher in clones containing the HSP27/UCOE combination compared to the clone containing only UCOE elements or to the control HEK293 cells. These results correlate with the MCP analyses, which showed that overexpression of HSP27 decreased the expression of Bax, caspase 3, cytochrome C, Beclin, and LC3II mRNA. Finally, this study suggests the potential utility of a cell engineering approach based on the overexpression of the human HSP27 protein for enhancing the production of recombinant viruses and proteins in HEK293 cells.