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Enhancement of human erythropoietin production in Chinese hamsterovary cells through supplementation of 30Kc19-30Kc6 fusion protein

Cited 7 time in Web of Science Cited 7 time in Scopus
Authors
Park, Hee Ho; Choi, Jeongseon; Lee, Hong Jai; Ryu, Jina; Park, Ju Hyun; Rhee, Won Jong; Park, Tai Hyun
Issue Date
2015
Publisher
Elsevier
Citation
Process Biochemistry, vol.50, pp. 973-980
Keywords
30Kc19-30Kc6 proteinSoluble expressionAnti-apoptosisErythropoietin (EPO)Productivity
Abstract
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.
Language
English
URI
http://hdl.handle.net/10371/95455
DOI
https://doi.org/10.1016/j.procbio.2015.03.008
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College of Engineering/Engineering Practice School (공과대학/대학원)Dept. of Chemical and Biological Engineering (화학생물공학부)Journal Papers (저널논문_화학생물공학부)
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