Enhancement of transient erythropoietin protein expression by valproic acid in CHO‐K1 suspension adapted cells

https://doi.org/10.22146/ijbiotech.52621

Yana Rubiyana(1*), Retno Damajanti Soejoedono(2), Adi Santoso(3)

(1) Department Biotechnology, IPB University, Bogor 16680, Indonesia; Research Center for Biotechnology, Indonesian Institute of Sciences
(2) Department of Animal Diseases and Veterinary Public Health, IPB University, Bogor 16680, Indonesia
(3) Research Center for Biotechnology, Indonesian Institute of Sciences, Cibinong 16911, Indonesia
(*) Corresponding Author

Abstract


Erythropoietin (EPO) is a therapeutic protein that is widely used to increase red blood cell production in chronic kidney failure. EPO protein can be produced quickly with a transient gene expression system (TGE). However, the titer produced using TGE is usually lower than the stable gene expression system (SGE). It has been known that TGE can be improved by histone deacetylase inhibitors (iHDACs) such as valproic acid (VPA). This study was conducted to examine the VPA effect on EPO protein expression in CHO‐K1 suspension adapted cells and to find the optimum concentration of VPA on transient EPO protein production. EPO proteins was quantified using the enzyme‐linked immunosorbent assay (ELISA) method. The optimization of VPA concentrations showed that VPA increased the EPO protein yield by up to 2‐fold in transient EPO production, and the optimum concentration of VPA was 4 mM. VPA optimization was very helpful to obtain the maximum increase in the transiently expressed protein. Furthermore, this study can be used as a model to produce EPO proteins or other recombinant proteins rapidly with TGE of CHO‐K1 suspension adapted cells.

Keywords


CHO‐K1; erythropoietin (EPO); transient gene expression system (TGE); histone deacetylase inhibitor (iHDAC); val‐ proic acid (VPA)

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References

Allen MJ, Boyce JP, Trentalange MT, Treiber DL, Rasmussen B, Tillotson B, Davis R, Reddy P. 2008. Identification of novel small molecule enhancers of protein production by cultured mammalian cells. Biotechnol Bioeng. 100(6):1193–1204. doi:10.1002/bit.21839.

Ashley RA, Dubuque SH, Dvorak B, Woodward SS, Williams SK, Kling PJ. 2002. Erythropoietin stimulates vasculogenesis in neonatal rat mesenteric microvascular endothelial cells. Pediatr Res. 51(4):472. doi:10.1203/00006450­200204000­00012.

Backliwal G, Hildinger M, Kuettel I, Delegrange F, Hacker DL, Wurm FM. 2008. Valproic acid: a viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures. Biotechnol Bioeng. 101(1):182–189. doi:10.1002/bit.21882.

Baldi L, Hacker DL, Adam M, Wurm FM. 2007. Recombinant protein production by large­scale transient gene expression in mammalian cells: state of the art and future perspectives. Biotechnol Lett. 29(5):677–684. doi:10.1007/s10529­006­9297­y.

Carpentier E, Paris S, Kamen AA, Durocher Y. 2007. Limiting factors governing protein expression following polyethylenimine­mediated gene transfer in HEK293­EBNA1 cells. J Biotechnol. 128(2):268– 280. doi:10.1016/j.jbiotec.2006.10.014.

Casadevall N, Durieux P, Dubois S, Hemery F, Lepage E, Quarré MC, Damaj G, Giraudier S, Guerci A, Laurent G. 2004. Health, economic, and quality­oflife effects of erythropoietin and granulocyte colonystimulating factor for the treatment of myelodysplastic syndromes: a randomized, controlled trial. Blood. 104(2):321–327. doi:10.1182/blood­2003­07­2252.

Egrie JC, Browne JK. 2001. Development and characterization of novel erythropoiesis stimulating protein (NESP). Br J Cancer. 84(1):3–13. doi:10.1054/bjoc.2001.1746.

Fan S, Maguire CA, Ramirez SH, Bradel­Tretheway B, Sapinoro R, Sui Z, Chakraborty­Sett S, Dewhurst S. 2005. Valproic acid enhances gene expression from viral gene transfer vectors. J Virol Methods. 125(1):23–33. doi:10.1016/j.jviromet.2004.11.023.

Fried W. 2009. Erythropoietin and erythropoiesis. Exp Hematol. 37(9):1007–1015. doi:10.1016/j.exphem.2009.05.010. Geisse S. 2009. Reflections on more than 10 years of TGE approaches. Protein Expression Purif. 64(2):99–107. doi:10.1016/j.pep.2008.10.017.

Haberland M, Montgomery RL, Olson EN. 2009. The many roles of histone deacetylases in development and physiology: implications for disease and therapy. Nat Rev Genet. 10(1):32. doi:10.1038/nrg2485.

Lechardeur D, Lukacs GL. 2006. Nucleocytoplasmic transport of plasmid DNA: a perilous journey from the cytoplasm to the nucleus. Hum Gene Ther. 17(9):882–889. doi:10.1089/hum.2006.17.882.

Marchion DC, Bicaku E, Daud AI, Sullivan DM, Munster PN. 2005. Valproic acid alters chromatin structure by regulation of chromatin modulation proteins. Cancer Res. 65(9):3815–3822. doi:10.1158/0008­ 5472.CAN­04­2478.

Michaelis M, Michaelis UR, Fleming I, Suhan T, Cinatl J, Blaheta RA, Hoffmann K, Kotchetkov R, Busse R, Nau H. 2004. Valproic acid inhibits angiogenesis in vitro and in vivo. Mol Pharmacol. 65(3):520–527. doi:10.1124/mol.65.3.520.

Pham PL, Kamen A, Durocher Y. 2006. Large­scale transfection of mammalian cells for the fast production of recombinant protein. Mol Biotechnol. 34(2):225– 237. doi:10.1385/MB:34:2:225.

Sami S, Höti N, Xu HM, Shen Z, Huang X. 2008. Valproic acid inhibits the growth of cervical cancer both in vitro and in vivo. J Biochem. 144(3):357–362. doi:10.1093/jb/mvn074.

Santoso A, Septisetyani EP, Meiyanto E, Ningrum RA. 2014. Expression of modified recombinant human erythropoietin in CHO­K1 cells and its in vitro proliferation assay in TF­1 cells. Indones J Pharm. 25(1):9– 16. doi:10.14499/indonesianjpharm25iss1pp9.

Sinhadri BCS. 2009. Effect of valproic acid on transient protein expression in HEK 293E suspension adapted cells. PhD Thesis, École Polytechnique Fédérale de Lausanne, Lausanne. Stein RS. 2003. The role of erythropoietin in the anemia of myelodysplastic syndrome. Clin Lymphoma. 4:S36– S40. doi:10.3816/CLM.2003.s.007.

Watson JD, Baker T, Gann A, Levine M, Losik R. 2014. Molecular biology of the gene. Boston: Pearson/CSH press. Wisnuwardhani PH, Septisetyani EP, Santoso A. 2017. Sequential Adaptation in Mammalian CHO­K1 Cells Producing Human Erythropoietin. Ann Bogor. 21(1):15–20. doi:10.14203/ann.bogor.2017.v21.n1.15­20.

Wulhfard S, Baldi L, Hacker DL, Wurm F. 2010. Valproic acid enhances recombinant mRNA and protein levels in transiently transfected Chinese hamster ovary cells. J Biotechnol. 148(2­3):128–132. doi:10.1016/j.jbiotec.2010.05.003.

Wulhfard S, Tissot S, Bouchet S, Cevey J, De Jesus M, Hacker DL, Wurm FM. 2008. Mild hypothermia improves transient gene expression yields several fold in Chinese hamster ovary cells. Biotechnol Prog. 24(2):458–465. doi:10.1021/bp070286c.

Yin H, Blanchard K. 2000. DNA methylation represses the expression of the human erythropoietin gene by two different mechanisms. Blood. 95(1):111–119. doi:10.1182/blood.V95.1.111.001k20_111_119.



DOI: https://doi.org/10.22146/ijbiotech.52621

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