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Open Access Research article

Optimization of heterologous protein production in Chinese hamster ovary cells under overexpression of spliced form of human X-box binding protein

Galina Gulis*, Kelly Cristina Rodrigues Simi, Renata Rodrigues de Toledo, Andrea Queiroz Maranhao and Marcelo Macedo Brigido

Author Affiliations

Institute of Biological Sciences, Department of Cell Biology, University of Brasilia, Campus Universitário Darcy Ribeiro, Brasília, DF 70910-900, Brazil

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BMC Biotechnology 2014, 14:26  doi:10.1186/1472-6750-14-26

Published: 11 April 2014

Abstract

Background

The optimization of protein production is a complex and challenging problem in biotechnology. Different techniques for transcription, translation engineering and the optimization of cell culture conditions have been used to improve protein secretion, but there remain many open problems involving post-translational modifications of the secreted protein and cell line stability.

Results

In this work, we focus on the regulation of secreted protein specific productivity (using a recombinant human immunoglobulin G (IgG)) by controlling the expression of the spliced form of human X-box binding protein (XBP-(s)) in Chinese hamster ovary cells (CHO-K1) under doxycycline (DOX) induction at different temperatures. We observed a four-fold increase in specific IgG productivity by CHO cells under elevated concentrations of DOX at 30°C compared to 37°C, without detectable differences in binding activity in vitro or changes in the structural integrity of IgG. In addition, we found a correlation between the overexpression of human XBP-1(s) (and, as a consequence, endoplasmic reticulum (ER) size expansion) and the specific IgG productivity under DOX induction.

Conclusions

Our data suggest the T-REx system overexpressing human XBP-1(s) can be successfully used in CHO-K1 cells for human immunoglobulin production.

Keywords:
CHO cells; Heterologous protein production; X-box binding protein; T-REx™ system; Doxycycline