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Transient and stable coagulation factor vii expression in a human cell line adapted to serum-free suspension culture

Grant number: 15/10675-0
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): March 13, 2016
Effective date (End): October 16, 2016
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Kamilla Swiech Antonietto
Grantee:Rafael Tagé Biaggio
Supervisor abroad: Amine A. Kamen
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Local de pesquisa : McGill University, Montreal, Canada  
Associated to the scholarship:14/01404-0 - Evaluation of growth and recombinant protein production of human cell lines adapted to serum-free suspension cultures, BP.DR

Abstract

Human coagulation factor VII (FVII) is a vitamin K-dependent glycoprotein that plays an important role in the blood coagulation process, being one of the key molecules involved in blood clot formation. Currently, the commercial product is produced in a murine cell line, BHK-21, which gives disadvantages considering the complexity of post-translational modifications of this protein. Murine cells simply do not possess the "machinery" required for human-type glycosylation. These differences in glycosylation pattern may be highly immunogenic in humans, and/or may be rapidly cleared the recombinant protein from circulation. Nevertheless, bioprocesses for recombinant protein production using mammalian cells have now focused on the use of serum-free suspension cultures. Based on this, the aim of this project is to express the coagulation factor VII (FVII) in a human Hek293 cell line adapted to serum-free suspension culture by stable and transient transfection. The technology learned during this project will be applied to the cells that are being used in the doctoral project.