title: Efficient expression of stable recombinant human insulin-like growth factor-1 fusion with human serum albumin in Chinese hamster ovary cells.
abstract: 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.
authors.affiliation: a The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology , Jiangnan University , Wuxi , China.
authors.forename: Aini
authors.initials: A
authors.lastname: Wan
authors.affiliation: b Laboratory of Molecular Pharmacology, School of Pharmaceutical Sciences , Jiangnan University , Wuxi , China.
authors.forename: Dongsheng
authors.initials: D
authors.lastname: Xu
authors.affiliation: b Laboratory of Molecular Pharmacology, School of Pharmaceutical Sciences , Jiangnan University , Wuxi , China.
authors.forename: Kedong
authors.initials: K
authors.lastname: Liu
authors.affiliation: a The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology , Jiangnan University , Wuxi , China.
authors.forename: Lin
authors.identifier: http://orcid.org/0000-0002-5708-4116
authors.initials: L
authors.lastname: Peng
authors.affiliation: b Laboratory of Molecular Pharmacology, School of Pharmaceutical Sciences , Jiangnan University , Wuxi , China.
authors.forename: Yanfei
authors.initials: Y
authors.lastname: Cai
authors.affiliation: b Laboratory of Molecular Pharmacology, School of Pharmaceutical Sciences , Jiangnan University , Wuxi , China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: c Jiangsu Institute of Hematology , The First Affiliated Hospital of Soochow University , Suzhou , China.
authors.forename: Yang
authors.initials: Y
authors.lastname: He
authors.affiliation: d Cyrus Tang Hematology Center and Ministry of Education Engineering Center of Hematological Disease , Soochow University , Suzhou , China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Yang
authors.affiliation: b Laboratory of Molecular Pharmacology, School of Pharmaceutical Sciences , Jiangnan University , Wuxi , China.
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
authors.affiliation: a The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology , Jiangnan University , Wuxi , China.
authors.forename: Huazhong
authors.initials: H
authors.lastname: Li
mesh_terms: D000818:Animals; D016466:CHO Cells; D049109:Cell Proliferation; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012709:Serum Albumin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D007334:Insulin-Like Growth Factor I
keywords: Bioactivity; Chinese hamster ovary cells; human serum albumin; insulin-like growth factor-1