Publications by authors named "Yushun Fu"
Improvement strategies for transient gene expression in mammalian cells.
Appl Microbiol Biotechnol
October 2024
Mammalian cells are suitable hosts for producing recombinant therapeutic proteins, with Chinese hamster ovary (CHO) and human embryonic kidney 293 (HEK293) cells being the most commonly used cell lines. Mammalian cell expression system includes stable and transient gene expression (TGE) system, with the TGE system having the advantages of short cycles and simple operation. By optimizing the TGE system, the expression of recombinant proteins has been significantly improved.
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- A transient gene expression system using CHO cells is crucial for quickly producing recombinant proteins, but low productivity is a major challenge.
- The study explores how co-overexpressing protein disulfide isomerase (PDI) and spliced X-box binding protein 1 (XBP-1s), combined with mild hypothermia, can enhance protein yields.
- Results showed that this combination significantly increased protein production and cell-specific productivity without affecting cell growth or viability, achieving substantial improvements in the expression of Adalimumab.
Previous work has shown that the EF-1α promoter of episomal vectors maintains high-level transgene expression in stably transfected Chinese hamster ovary (CHO) cells. However, the transgene expression levels need to be further increased. Here, we first incorporated matrix attachment regions (MARs), ubiquitous chromatin opening element (UCOE), stabilizing anti repressor elements 40 (STAR 40) elements into episomal vector at different sites and orientations, and systemically assessed their effects on transgene expression in transfected CHO-K1 cells.
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