Adaptation process of engineered cell line FCHO/IL-24 stably secreted rhIL-24 in serum-free suspension culture
文献类型：期刊论文
| 作者 | Hua, Jilei1; Wei, Yuexian1; Zhang, Yao1,3; Xu, Hanli1; Ge, Jianlin1; Liu, Mengzhe1; Wang, Yuqi1; Shi, Yinan1,2; Hou, Lingling1; Jiang, Hong1 |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2022-11-01 |
| 卷号 | 199页码:10 |
| ISSN号 | 1046-5928 |
| 关键词 | The recombinant human IL-24 protein Engineered FCHO IL-24 cell line Serum-free culture Suspension culture CHO cells |
| DOI | 10.1016/j.pep.2022.106154 |
| 英文摘要 | Interleukin-24 (IL-24) displays tumor cell-specific proliferation inhibition in vitro and in vivo. Recombinant human IL-24 (rhIL-24) has significantly higher activity, yet significantly lower expression level in mammalian cells than in bacteria. To further realize therapeutic potential of IL-24, we enhanced rhIL-24 expression in mammalian cell systems by adapting engineered Flp-InTMCHO/IL-24 (FCHO/IL-24) cells (adherent cultured in Ham's F12 medium with 10% serum) to serum-free suspension culture. First, MTT assay showed that among four different media (F12, DMEM/F12, 1640 and DMEM), DMEM/F12 medium was the most suitable media for lower-serum adherent culture. Then, cells were adherently cultured in DMEM/F12 with serum concentration reduced from 10% to 0.5% in a gradient manner. Compared to cells in 10% serum, cells in 0.5% serum displayed significantly lower relative cell viability by 40%, increased G0/G1 phase arrest (8.5 & PLUSMN; 2.4%, p < 0.05), decreased supernatant rhIL-24 concentration by 73%, and altered metabolite profiles, such as glucose, lactate and ammonia concentration. Next, the cells were directly adapted to 0.5% serum suspension culture in 125 mL shake flask at 119 rpm with the optimal cell seeding density of 5 x 105 cells/mL (3.3 times higher than that of adherent culture), under which the concentration of rhIL-24 in culture medium was stable at 3.5 ng/mL. Finally, cells adapted to 0.5% serum proliferated better in serum-free medium EdenTM-B300S with higher rhIL-24 expression level compared to CDM4CHO. The successful adaptation of engineered cells FCHO/IL-24 laid foundation for adapting cells from adherent culture to suspension serum-free culture to mass produce rhIL-24 protein for therapeutic purposes. |
| WOS关键词 | CHO-CELLS ; GROWTH ; PROLIFERATION ; PRODUCTIVITY ; METABOLISM ; MDA-7 ; GENE |
| 资助项目 | Beijing Natural Science Foun-dation[7214242] ; Fundamental Research Funds for the Central Universities[2021RC207] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| WOS记录号 | WOS:000851336400003 |
| 资助机构 | Beijing Natural Science Foun-dation ; Fundamental Research Funds for the Central Universities |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/54638]  |
| 专题 | 中国科学院过程工程研究所 |
| 通讯作者 | Hou, Lingling; Jiang, Hong |
| 作者单位 | 1.Beijing Jiaotong Univ, Coll Life Sci & Bioengn, 3 Shangyuancun, Beijing 100044, Peoples R China 2.China Acad Chinese Med Sci, Inst Acupuncture & Moxibust, Beijing 100700, Peoples R China 3.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China |
| 推荐引用方式 GB/T 7714 | Hua, Jilei,Wei, Yuexian,Zhang, Yao,et al. Adaptation process of engineered cell line FCHO/IL-24 stably secreted rhIL-24 in serum-free suspension culture[J]. PROTEIN EXPRESSION AND PURIFICATION,2022,199:10. |
| APA | Hua, Jilei.,Wei, Yuexian.,Zhang, Yao.,Xu, Hanli.,Ge, Jianlin.,...&Jiang, Hong.(2022).Adaptation process of engineered cell line FCHO/IL-24 stably secreted rhIL-24 in serum-free suspension culture.PROTEIN EXPRESSION AND PURIFICATION,199,10. |
| MLA | Hua, Jilei,et al."Adaptation process of engineered cell line FCHO/IL-24 stably secreted rhIL-24 in serum-free suspension culture".PROTEIN EXPRESSION AND PURIFICATION 199(2022):10. |
入库方式： OAI收割
来源：过程工程研究所
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