Optimization of heterologous protein production in Chinese hamster ovary cells under overexpression of spliced form of human X-box binding protein
| Autor: | Renata Rodrigues de Toledo, Kelly Cristina Rodrigues Simi, Marcelo M. Brigido, Andrea Queiroz Maranhão, Galina Gulis |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
X-Box Binding Protein 1
Heterologous Regulatory Factor X Transcription Factors Biology Endoplasmic Reticulum Transfection Retinoblastoma-like protein 1 Cricetulus Protein biosynthesis Animals Humans Protein Isoforms T-REx™ system Transcription factor Heterologous protein production Binding protein Chinese hamster ovary cell Temperature CHO cells Molecular biology Culture Media Cell biology DNA-Binding Proteins EIF4EBP1 Cell culture Doxycycline Immunoglobulin G Proteínas Research Article X-box binding protein Plasmids Transcription Factors Biotechnology |
| Zdroj: | Repositório Institucional da UnB Universidade de Brasília (UnB) instacron:UNB BMC Biotechnology |
| ISSN: | 1472-6750 |
| DOI: | 10.1186/1472-6750-14-26 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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