Advanced search
Start date
Betweenand
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Human cell lines: A promising alternative for recombinant FIX production

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