S-Space College of Engineering/Engineering Practice School (공과대학/대학원) Dept. of Chemical and Biological Engineering (화학생물공학부) Journal Papers (저널논문_화학생물공학부)
Enhancement of human erythropoietin production in Chinese hamsterovary cells through supplementation of 30Kc19-30Kc6 fusion protein
- Park, Hee Ho; Choi, Jeongseon; Lee, Hong Jai; Ryu, Jina; Park, Ju Hyun; Rhee, Won Jong; Park, Tai Hyun
- Issue Date
- Process Biochemistry, vol.50, pp. 973-980
- 30Kc19-30Kc6 protein; Soluble expression; Anti-apoptosis; Erythropoietin (EPO); Productivity
- tThe importance of anti-apoptosis in mammalian cell culture has been widely recognized. We reportedpreviously that expression of Bombyx mori 30 K genes in Chinese hamster ovary (CHO) cells increasesrecombinant protein production by inhibiting apoptosis and enhancing specific productivity. However,previous studies have shown expression of the anti-apoptotic 30Kc6 protein as inclusion bodies inEscherichia coli. 30Kc19 protein, another silkworm hemolymph protein, has cell-penetrating and recom-binant protein productivity-improving properties, and we found that it improves soluble expressionof its partner. In this study, we fused 30Kc6 with 30Kc19 as an expression partner to express a sol-uble fused protein and to deliver the protein to cells. Supplementing the recombinant 30Kc19-30Kc6fusion protein in cell culture medium increased viability, effectively penetrated the cells, and inhibitedCHO cell apoptosis by 68%. Moreover, the mitochondrial membrane potential and ATP generation alsoincreased by 50% and 33%, respectively. Erythropoietin (EPO) productivity increased by > 30% because ofthe anti-apoptotic effect and increased specific productivity. These results demonstrate the potential useof this fusion protein as a supplement in mammalian cell culture during production of biopharmaceuticalproteins.
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