Efficient expression of stable recombinant human insulin-like growth factor-1 fusion with human serum albumin in Chinese hamster ovary cells
| Autor: | Aini Wan, Jian Jin, Kedong Liu, Yanfei Cai, Huazhong Li, Dongsheng Xu, Yang He, Jianfeng Yang, Yun Chen, Lin Peng |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Recombinant Fusion Proteins Gene Expression CHO Cells Biology Biochemistry Mice 03 medical and health sciences Cricetulus Cricetinae medicine Animals Humans Cloning Molecular Insulin-Like Growth Factor I Serum Albumin Cell Proliferation Chinese hamster ovary cell General Medicine Metabolism Human serum albumin Molecular biology Recombinant human insulin-like growth factor 1 030104 developmental biology NIH 3T3 Cells Plasmids Biotechnology medicine.drug |
| Zdroj: | Preparative Biochemistry & Biotechnology. 47:678-686 |
| ISSN: | 1532-2297 1082-6068 |
| DOI: | 10.1080/10826068.2017.1303612 |
| Popis: | Insulin-like growth factor-1 (IGF-1) plays a crucial role in cell development, differentiation, and metabolism, and has been a potential therapeutic agent for many diseases. Chinese hamster ovary (CHO) cells are widely used for production of recombinant therapeutic proteins, but the expression level of IGF-1 in CHO cells is very low (1,500 µg/L) and the half-life of IGF-1 in blood circulation is only 4.5 min according to previous studies. Therefore, IGF-1 was fused to long-circulating serum protein human serum albumin (HSA) and expressed in CHO cells. After 8-day fed-batch culture, the expression level of HSA-IGF-1 reached 100 mg/L. The fusion protein HSA-IGF-1 was purified with a recovery of 35% using a two-step chromatographic procedure. According to bioactivity assay, the purified HSA-IGF-1 could stimulate the proliferation of NIH3T3 cells in a dose-dependent fashion and promote the cell-cycle progression. Besides this, HSA-IGF-1 could bind to IGF-1 receptor on cell membrane and activate the intracellular PI3K/AKT signaling pathway. Our study suggested that HSA fusion technology carried out in CHO cells not only provided bioactivity in HSA-IGF-1 for further research but also offered a beneficial strategy to produce other similar cytokines in CHO cells. |
| Databáze: | OpenAIRE |
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