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

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Costa e Silva, Luciano ; Santos, Matheus Henrique ; Heluy, Tarik Reis ; Biaggio, Rafael Tage ; Ferreira, Alexander Rodrigo ; Milhomens, Jonathan ; Kashima, Simone ; El Nemer, Wassim ; El Hoss, Sara ; Picanco-Castro, Virginia ; Covas, Dimas Tadeu ; Swiech, Kamilla

• Tipo de documento: Artigo Científico
• Fonte: ENGINEERING REPORTS; v. 2, n. 6, p. 16-pg., 2020-06-01.

Resumo

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 mu g/mL, 112.7 mu g/mL, and 571 mu g/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. (AU)

• Processo FAPESP: 12/04629-8 - Estabelecimento de uma plataforma de produção de proteínas recombinantes terapêuticas em células humanas
• Beneficiário: Kamilla Swiech Antonietto
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 13/08135-2 - CTC - Centro de Terapia Celular
• Beneficiário: Dimas Tadeu Covas
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs