Pfizer, 1 Burtt Rd, Andover, MA 01810, USA.
Biotechnol Lett. 2011 Sep;33(9):1773-9. doi: 10.1007/s10529-011-0628-2. Epub 2011 May 6.
The Chinese hamster ovary (CHO) cell line is one of the most widely used mammalian cell lines for biopharmaceutical production. We have developed and characterized a gene expression microarray (WyeHamster2a) specific for CHO cells that has enabled the study of ~3,500 sequences. Analysis of multiple sets of replicate scans showed that data derived from the WyeHamster2a array is highly reproducible confirming it as a robust tool for profiling. Twelve gene sequences were selected for follow-up RT-qPCR to confirm the accuracy and precision of the microarray results. In all but the most subtle gene expression differences, the microarray proved to be a reliable measure of differential gene expression. Finally, we were able to quantify the difference between using a bona fide CHO-specific microarray for profiling CHO cells versus an alternate, commercially available, rodent microarray such as a mouse or rat-specific format.
中国仓鼠卵巢(CHO)细胞系是用于生物制药生产的最广泛使用的哺乳动物细胞系之一。我们开发并鉴定了一种针对 CHO 细胞的基因表达微阵列(WyeHamster2a),该微阵列可用于研究约 3500 个序列。对多组重复扫描的分析表明,WyeHamster2a 阵列获得的数据具有高度可重复性,证实了它作为一种用于分析的强大工具。选择了 12 个基因序列进行后续 RT-qPCR,以确认微阵列结果的准确性和精密度。在除了最细微的基因表达差异外,微阵列都被证明是一种可靠的差异基因表达衡量标准。最后,我们能够量化使用真正的 CHO 特异性微阵列进行 CHO 细胞分析与使用替代的、市售的、啮齿动物微阵列(如小鼠或大鼠特异性格式)之间的差异。