Serum‐free suspension cultured human cells can produce a high‐level of recombinant human erythropoietin

Abstract

Abstract Currently, the majority of commercial recombinant therapeutic proteins are produced in nonhuman expression systems. Human cells are capable of producing recombinant proteins with post‐translational modifications that are more similar to native proteins which reduce immunogenicity. This article describes the potential of the human cell lines Sk‐Hep‐1, HKB‐11, and Huh‐7, cultured in serum‐free suspension conditions, to produce a complex recombinant glycoprotein, human erythropoietin (EPO). Recombinant cell lines were generated by lentiviral transduction and sorted by flow cytometry. All the recombinant cells presented high‐specific cell growth rates and a high level of rhEPO production. Maximum rhEPO concentrations achieved by Sk‐Hep‐1, HKB‐11, and Huh‐7 cells were 112.5 μg/mL, 112.7 μg/mL, and 571 μg/mL, respectively. The levels of rhEPO production by Huh‐7 cells were higher than the levels commonly reported in the literature for serum‐free suspension cultures. The rhEPO produced demonstrated biological activity measured through in vitro differentiation of CD34+ cells. In view of this, we demonstrate that Sk‐Hep‐1, HKB‐11, and Huh‐7 cell lines have good characteristics to be used as host cells for the production of complex recombinant glycoproteins, with special emphasis on Huh‐7 due to enhanced EPO production.

Keywords

• erythropoietin
• human cell lines
• recombinant glycoproteins
• serum‐free suspension culture