Effects of TCA Cycle Intermediates on hCTLA4Ig Production

초록

Plant-made pharmaceuticals have received considerable interest since they do not have risk of harboring animal-derived pathogens and can produce at much lower cost in plant cells than that in mammalian cells. Transgenic plant cell culture system with the signal peptide for the secretion of target protein into the extracellular medium and the inducible RAmy3D promoter was used for the production of the recombinant therapeutic protein in order to overcome the constraint of low production level. However the RAmy3D promoter system has a drawback in decreasing cell viability because the promoter is induced only by sugar starvation. In this study, improvement of hCTLA4Ig productivity using transgenic cell line was investigated in flask cultures by supplementing alternative energy sources. To increase recombinant protein production, pyruvate and amino acids have been already used as alternative energy sources in mammalian cells. Two forms of fumarate, which is a TCA cycle intermediate, was used as a new energy source to increase viability and productivity. Supplementation of 40 mM fumarate acid in production medium enhanced the productivity by 1.2 fold compared to those of control and fumarate sodium salt form.