Efficient production of recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium

Abstract

A fructose-based cell culture is suitable for the process control of protein production because of slow sugar consumption rate and low lactate accumulation. The fructose transporter, GLUT5, mediates its incorporation into cells and is required for the fructosebased culture. In order to produce efficiently recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium, an IgG and GLUT5 co-expression vector was constructed and transfected into the human myeloma derived cell line, SC-01MFP, which produced stably recombinant proteins. The cell proliferation in the fructose-based medium was improved by the GLUT5 gene transfection. The recombinant IgG production of the cells cultured in the fructose-based medium exhibited about two-fold increase of that in the glucose-based medium. Flow cytometoric analysis indicated that the GLUT5 protein expression level in cell surface was increased in the fructose-based medium. An exogenous but not endogenous GLUT5 transcription activator remarkably raised IgG productivity in the fructose-based medium when compared to that in the glucose-based medium, suggesting that exogenous GLUT5 expression may be involved in it. The GLUT5 co-expression system may be useful for efficient production of recombinant proteins by the fructose-based cell culture.

Knockout of the caspase 8‐associated protein 2 gene improves recombinant protein expression in HEK293 cells through up‐regulation of the cyclin‐dependent kinase inhibitor 2A gene

“…The gene targets for cell modification are usually selected based on their known biological functions and the desired phenotypic outcome. For example, proapoptotic and antiapoptotic genes have been manipulated to extend the longevity of cultured cells (Baek et al, 2017; Cost et al, 2010); and genes involved in sugar transport have been manipulated to improve the metabolic efficiency of the producing cells (Inoue et al, 2010; Paredes et al, 1999). However, this approach is limited to genes with known function, whose altered expression may achieve the desired phenotype.…”

Comparative Metabolic Analysis of CHO Cell Clones Obtained through Cell Engineering, for IgG Productivity, Growth and Cell Longevity

“…These engineered cells were characterized by a better use of the main carbon source, consuming a lower amount of carbon molecules and producing less lactate. To further investigate the impact of SLC2A5 gene overexpression, in 2010 Inoue and collaborators reported that cells derived from human myeloma overexpressing GLUT5 were able to achieve more than 1.5 times the cell density reached by wild-type cells and produce more than 2 times the amount of recombinant protein [ 14 ].…”

Nucleofection of whole murine retinas

“…Ning et al 2012). The expression of recombinant proteins under the control of a cell-specific promoter allows the characterization of their role in a cell-specific manner (Montana et al 2011;Cai et al 2010), or the production of promising therapeutic proteins such as immunoglobulins (Inoue et al 2010). Although the potential of transfection is enormous, transfection of many primary cells present many technical challenges.…”