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Evaluation of growth and recombinant protein production of human cell lines adapted to serum-free suspension cultures

Grant number: 14/01404-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2014
Effective date (End): October 05, 2018
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Kamilla Swiech Antonietto
Grantee:Rafael Tagé Biaggio
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:12/04629-8 - Establishment of a production platform for recombinant therapeutic proteins in human cells, AP.JP
Associated scholarship(s):15/10675-0 - Transient and stable coagulation factor vii expression in a human cell line adapted to serum-free suspension culture, BE.EP.DR

Abstract

Mammalian cell lines are the most used system for the production of recombinant therapeutic proteins, since they are capable of performing post-translational modifications and protein folding in an authentic manner, thus producing recombinant protein with appropriate quality. Due to the growing demand for these products and the high cost and complexity of manufacturing processes, biopharmaceutical companies are interested in developing optimized biological processes, able to generate high levels of recombinant proteins with therapeutic quality. Human cell lines have attracted much interest because they are able to produce glycosylated proteins more similar to native human proteins, avoiding potential immunogenic responses. However, they are not properly characterized in terms of a future commercial application. Based on this, the main objective of this project is to evaluate the growth and production of recombinant proteins of SK-Hep-1, HKB-11 and Huh-7 human cell lines adapted to grow in suspension serum-free cultures in order to establish a platform for the production of therapeutic recombinant proteins based on these promising human cell lines. The previously adapted SK-Hep-1 and HKB-11 human cell lines and the Huh-7 human cell line, the latter will be subsequently adapted to serum-free suspension culture and all of the three cell lines will be cultured in bioreactors. The kinetic parameters of growth and metabolism of the cells will be analyzed. The potential for recombinant Factor VII (FVII ) production in the same adapted cell lines will also be evaluated. The results of growth and yield of cell lines (FVII) will be compared with the commercial HEK293 human cell line also adapted for growth in suspension and serum-free medium.