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All Journal Indonesian Journal of Cancer Chemoprevention
Endah Puji Septisetyani
Research Center for Biotechnology, Indonesian Institute of Sciences, Cibinong, Bogor 16911
Biochemistry, Genetics & Molecular Biology Medicine & Pharmacology

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Expression of Human Erythropoietin Containing 2 Additional N-Link in CHO-K1 Cells under Different Culture Conditions Adi Santoso; Larasati Larasati; Arizah Kusumawati; Popi Hadi Wisnuwardhani; Ratih Asma Ningrum; Endah Puji Septisetyani
Indonesian Journal of Cancer Chemoprevention Vol 10, No 1 (2019)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev10iss1pp7-15

Abstract

Human erythropoietin (hEPO) is a glycoprotein that regulates the formation of erythrocytes and mainly used in anemia patients. Previously, we have reported the expression of modified human EPO with 2 additional N-linked in mammalian cell CHO-K1. The aim of this current research was to study the optimum condition for modified recombinant hEPO (rhEPO) production in CHO-K1. To do this, several parameters of culture conditions were applied including antibiotic concentrations, seeding densities, time of incubations, fetal bovine serum (FBS) concentrations and cell culture media. The result showed that the presence of antibiotic G418 improved the expression level with the highest was at 1% of concentration. Meanwhile, seeding density of 2–3x105 cells/6 cm dish and seven day of incubation time were the best condition for rhEPO protein expression. From five different combination media used, F12 medium with 10% FBS gave the highest expression of rhEPO protein. From this study was also found that at passage 16 the expression level was still increasing proving that the clone expressing the protein of our interest is promisingly stable.Keywords : EPO, erythropoietin, protein expression, CHO-K1, optimation
Co-Authors Adi Santoso Arizah Kusumawati Larasati Larasati Popi Hadi Wisnuwardhani Ratih Asma Ningrum