Post-translational modification profile of recombinant factor VII produced in huma...
| Full text | |
| Author(s): |
Bomfim, Aline de Sousa
[1, 2, 3]
;
Correa de Freitas, Marcela Cristina
[2, 3, 4]
;
Picanco-Castro, Virginia
[2, 3]
;
Soares Neto, Mario de Abreu
[2, 3]
;
Swiech, Kamilla
[2, 3, 5]
;
Covas, Dimas Tadeu
[2, 3, 4]
;
de Sousa Russo, Elisa Maria
[1, 2, 3]
Total Authors: 7
|
| Affiliation: | [1] Univ Sao Paulo, Dept Clin Toxicol & Food Sci Anal, Fac Pharmaceut Sci Ribeirao Preto, 2501 Tenente Catao Roxo St, BR-14051140 Ribeirao Preto, SP - Brazil
[2] Univ Sao Paulo, Ctr Cell Based Therapy, 2501 Tenente Catao Roxo St, BR-14051140 Ribeirao Preto, SP - Brazil
[3] Univ Sao Paulo, Reg Blood Ctr, 2501 Tenente Catao Roxo St, BR-14051140 Ribeirao Preto, SP - Brazil
[4] Univ Sao Paulo, Dept Clin Med, Fac Med Ribeirao Preto, 2501 Tenente Catao Roxo St, BR-14051140 Ribeirao Preto, SP - Brazil
[5] Univ Sao Paulo, Dept Pharmaceut Sci, Fac Pharmaceut Sci Ribeirao Preto, 2501 Tenente Catao Roxo St, BR-14051140 Ribeirao Preto, SP - Brazil
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | Protein Expression and Purification; v. 121, p. 149-156, MAY 2016. |
| Web of Science Citations: | 2 |
| Abstract | |
Factor IX (FIX) is a vitamin K-dependent protein, and it has become a valuable pharmaceutical in the Hemophilia B treatment. We evaluated the potential of recombinant human FIX (rhFIX) expression in 293T and SK-Hep-1 human cell lines. SK-Hep-1-FIX cells produced higher levels of biologically active protein. The growth profile of 293T-FIX cells was not influenced by lentiviral integration number into the cellular genome. SK-Hep-1-FIX cells showed a significantly lower growth rate than SK-Hep-1 cells. gamma-carboxylation process is significant to FIX biological activity, thus we performed a expression analysis of genes involved in this process. The 293T gene expression suggests that this cell line could efficiently carboxylate FIX, however only 28% of the total secreted protein is active. SK-Hep-1 cells did not express high amounts of VKORC1 and carboxylase, but this cell line secreted large amounts of active protein. Enrichment of culture medium with Ca+2 and Mg+2 ions did not affect positively rhFIX expression in SK-Hep-1 cells. In 293T cells, the addition of 0.5 mM Ca+2 and 1 mM Mg+2 resulted in higher rhFIX concentration. SK-Hep-1 cell line proved to be very effective in rhFIX production, and it can be used as a novel biotechnological platform for the production of recombinant proteins. (C) 2016 Elsevier Inc. All rights reserved. (AU) | |
| FAPESP's process: | 12/04629-8 - Establishment of a production platform for recombinant therapeutic proteins in human cells |
| Grantee: | Kamilla Swiech Antonietto |
| Support type: | Research Grants - Young Investigators Grants |