Production Of coagulation factor VII in human cell lines Sk-Hep-1 and HBK-11

Autor(es)

Correa de Freitas, Marcela Cristina ; Bomfim, Aline de Sousa ; Mizukami, Amanda ; Picanco-Castro, Virginia ; Swiech, Kamilla ; Covas, Dimas Tadeu

• Tipo de documento: Artigo Científico
• Fonte: Protein Expression and Purification; v. 137, p. 26-33, SEP 2017.
• Citações Web of Science: 1

Resumo

Recombinant factor VII (rFVII) is the main therapeutic choice for hemophilia patients who have developed inhibitory antibodies against conventional treatments (FVIII and FIX). Because of the post translational modifications, rFVII needs to be produced in mammalian cell lines. In this study, for the first time, we have shown efficient rFVII production in HepG2, Sk-Hep-1, and HKB-11 cell lines. Experiments in static conditions for a period of 96 h showed that HepG2-FVII produced the highest amounts of rhFVII, with an average of 1843 ng/mL. Sk-hep-1-FVII cells reached a maximum protein production of 1432 ng/mL and HKB-11-FVII cells reached 1468 ng/mL. Sk-Hep-1-rFVII and HKB-11-rFVII were selected for the first step of scale-up. Over 10 days of spinner flask culture, HKB-11 and SK-Hep-1 cells showed a cumulative production of rFVII of 15214 and 202.6 mu g in 50 mL, respectively. Thus, these human cell lines can be used for an efficient production of recombinant FVII. With more investment in basic research, human cell lines can be optimized for the commercial production of different bio therapeutic proteins. (C) 2017 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 11/01736-5 - Clonagem e expressão do fator VII de coagulação saguínea em linhagens celulares humanas
• Beneficiário: Marcela Cristina Corrêa de Freitas
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• 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
• Processo FAPESP: 16/02433-0 - Purificação, formulação e ensaio pré-clínico do FVIII recombinante produzido em células humanas.
• Beneficiário: Virginia Picanço e Castro
• Modalidade de apoio: Auxílio à Pesquisa - Regular