Efficient Interleukin-21 Production by Optimization of Codon and Signal Peptide in Chinese Hamster Ovarian Cells
| Autor: | Hee Jun Cho, Hee Gu Lee, Bo Yeon Kim, Jeewon Lim, Kyoung Eun Baek, Yo Sep Hwang, In Pyo Choi, Jong-Tae Kim, Byung Moo Oh, Sang Yoon Park |
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| Rok vydání: | 2018 |
| Předmět: |
Signal peptide
STAT3 Transcription Factor medicine.medical_treatment Genetic Vectors CHO Cells Protein Sorting Signals Protein Engineering Transfection Applied Microbiology and Biotechnology Azurocidin Interleukin 21 Interferon-gamma Immune system Cricetulus medicine Animals Humans Secretion Phosphorylation Codon Biosimilar Pharmaceuticals Chemistry Chinese hamster ovary cell Interleukins General Medicine Recombinant Proteins Cell biology Cytokine Biotechnology |
| Zdroj: | Journal of microbiology and biotechnology. 29(2) |
| ISSN: | 1738-8872 |
| Popis: | Interleukin-21 is a common γ-chain cytokine that controls the immune responses of B cells, T cells, and natural killer cells. Targeting IL-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. However, the practical application of IL-21 is limited by the high production cost. In this study, we improved IL-21 production by codon optimization and selection of appropriate signal peptide in CHO-K1 cells. Codon-optimized or non-optimized human IL-21 was stably transfected into CHO-K1 cells. IL-21 expression was 10-fold higher for codon-optimized than non-optimized IL-21. We fused five different signal peptides to codon-optimized mature IL-21 and evaluated their effect on IL-21 production. The best result (a 3-fold increase) was obtained using a signal peptide derived from human azurocidin. Furthermore, codon-optimized IL-21 containing the azurocidin signal peptide promoted IFN-γ secretion and STAT3 phosphorylation in NK-92 cells similar to codon-optimized IL-21 containing original signal peptide. Collectively, these results indicate that codon optimization and azurocidin signal peptides provide an efficient approach for the high-level production of IL-21 as a biopharmaceutical. |
| Databáze: | OpenAIRE |
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