Glycosylation and post-translational modification gene expression analysis by DNA microarrays for cultured mammalian cells
- 作者：Arthur Nathan Brodsky ; Mary Caldwell ; Sarah W. Harcum ; harcum@clemson.edu
- 关键词：CHO cells ; Chinese hamster ovary cells ; NS0 cells ; a mouse myeloma cell line ; real-time qRT-PCR ; real-time quantitative reverse transcription polymerase chain reaction ; PM ; perfect match ; MM ; mismatch
- 刊名：Methods
- 出版年：2012
- 期刊代码：103_10462023
- 类别：imm
- 出版时间：March, 2012
- 卷：56
- 期：3
- 页码：408-417
- 文件大小：728 K
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.