Enhanced recombinant M-CSF production in CHO cells by glycerol addition: model and validation

被引:0
作者
Chi-Hsien Liu
Li-Hsin Chen
机构
[1] Chang Gung University,Graduate Institute of Biochemical and Biomedical Engineering
来源
Cytotechnology | 2007年 / 54卷
关键词
CHO cells; Controlled proliferation; Glycerol; M-CSF; Model; Optimization;
D O I
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中图分类号
学科分类号
摘要
Addition of stimulatory chemical such as glycerol was found to increase recombinant protein production in Chinese hamster ovary (CHO) cells. However, glycerol influenced cell mitosis and reduced cell growth rate. We developed a controlled proliferation strategy to utilize the stimulation of glycerol on recombinant protein production and mitigate the problem of growth inhibition. The approach is to apply a two-stage process, where cells are cultured without glycerol for a period of time in order to obtain enough cell density and then glycerol is added to achieve high specific productivity. In addition, a model for predicting the profiles of cell proliferation and recombinant protein production was developed and validated. A two-stage process, addition of 1% glycerol after 1 day of growth, could increase the final production of macrophage-colony stimulating factor (M-CSF) by 38% compared with the value obtained without addition of glycerol.
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页码:89 / 96
页数:7
相关论文
共 37 条
[1]  
Arden N(2006)Regulating apoptosis in mammalian cell cultures Cytotechnology 50 77-92
[2]  
Betenbaugh AMJ(1996)Chemical chaperones correct the mutant phenotype of the delta F508 cystic fibrosis transmembrane conductance regulator protein Cell Stress Chaperones 1 117-125
[3]  
Brown CR(2004)Maximizing interferon-γ production by Chinese hamster ovary cells through temperature shift optimization: experimental and modeling Biotechnol Bioeng 85 177-184
[4]  
Hong-Brown LQ(1998)Rhodanese folding is controlled by the partitioning of its folding intermediates Biochim Biophys Acta 1382 120-128
[5]  
Biwersi J(2000)Role of protein-solvent interactions in refolding: effects of cosolvent additives on the renaturation of porcine pancreatic elastase at various pHs Arch Biochem Biophys 375 220-228
[6]  
Verkman AS(2005)Effects of supplementation of various medium components on Chinese hamster Cytotechnology 47 37-49
[7]  
Welch WJ(2002)Clone and expression of mutant M-CSF and its receptor from human leukemic cell line J6–1 Leukemia Res 26 377-382
[8]  
Fox SR(2005)Efficient refolding of aggregation-prone citrate synthase by polyol osmolytes: how well are protein folding and stability aspects coupled J Biol Chem 280 15553-15560
[9]  
Patel UA(2006)Reactor engineering in large scale animal cell culture Cytotechnology 50 9-33
[10]  
Yap MGS(2005)Enhanced production of monomeric interferon by CHO cells through the control of culture conditions Biotechnol Prog 21 22-30