Glycosylation and post-translational modification gene expression analysis by DNA microarrays for cultured mammalian cells

被引:6
作者
Brodsky, Arthur Nathan [1 ]
Caldwell, Mary [1 ]
Harcum, Sarah W. [1 ]
机构
[1] Clemson Univ, Dept Bioengn, Rhodes Res Ctr 301, Clemson, SC 29634 USA
关键词
DNA microarrays; Glycosylation; Chinese hamster ovary cells; NS0; cells; CHINESE-HAMSTER OVARY; RT-PCR METHOD; N-GLYCOSYLATION; CHO-CELLS; TRANSCRIPTIONAL ANALYSIS; RECOMBINANT PROTEINS; DISSOLVED-OXYGEN; SODIUM-BUTYRATE; MESSENGER-RNA; SYBR GREEN;
D O I
10.1016/j.ymeth.2011.10.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
DNA microarray analysis of gene expression has become a valuable tool for bioprocessing research aimed at improving therapeutic protein yields. The highly parallel nature of DNA microarray technology allows researchers to assess hundreds of gene simultaneously, essentially enabling genome-wide snapshots. The quality and amount of therapeutic proteins produced by cultured mammalian cells rely heavily on the culture environment. In order to implement beneficial changes to the culture environment, a better understanding of the relationship between the product quality and culture environment must be developed. By analyzing gene expression levels under various environmental conditions, light can be shed on the underlying mechanisms. This paper describes a method for evaluating gene expression changes for cultured NS0 cells, a mouse-derived myeloma cell line, under culture environment conditions, such as ammonia buildup, known to affect product quality. These procedures can be easily adapted to other environmental conditions and any mammalian cell lines cultured in suspension, so long as a sufficient number of gene sequences are publicly available. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:408 / 417
页数:10
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