| Full text | |
| Author(s): Show less - |
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
Total Authors: 12
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| Document type: | Journal article |
| Source: | ENGINEERING REPORTS; v. 2, n. 6, p. 16-pg., 2020-06-01. |
| 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 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) | |
| FAPESP's process: | 12/04629-8 - Establishment of a production platform for recombinant therapeutic proteins in human cells |
| Grantee: | Kamilla Swiech Antonietto |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 13/08135-2 - CTC - Center for Cell-Based Therapy |
| Grantee: | Dimas Tadeu Covas |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |