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通过靶向RNA测序检测中国仓鼠卵巢抗体产生细胞系中的突变

Mutation Detection in an Antibody-Producing Chinese Hamster Ovary Cell Line by Targeted RNA Sequencing.

作者信息

Zhang Siyan, Hughes Jason D, Murgolo Nicholas, Levitan Diane, Chen Janice, Liu Zhong, Shi Shuangping

机构信息

Biologics & Vaccines, Merck Research Laboratories, Kenilworth, NJ 07033, USA.

Biology & Genetics Informatics, Merck Research Labs IT, Merck & Co, Boston, MA 02115, USA.

出版信息

Biomed Res Int. 2016;2016:8356435. doi: 10.1155/2016/8356435. Epub 2016 Mar 20.

Abstract

Chinese hamster ovary (CHO) cells have been used widely in the pharmaceutical industry for production of biological therapeutics including monoclonal antibodies (mAb). The integrity of the gene of interest and the accuracy of the relay of genetic information impact product quality and patient safety. Here we employed next-generation sequencing, particularly RNA-seq, and developed a method to systematically analyze the mutation rate of the mRNA of CHO cell lines producing a mAb. The effect of an extended culturing period to mimic the scale of cell expansion in a manufacturing process and varying selection pressure in the cell culture were also closely examined.

摘要

中国仓鼠卵巢(CHO)细胞已在制药行业中广泛用于生产包括单克隆抗体(mAb)在内的生物治疗药物。目的基因的完整性和遗传信息传递的准确性会影响产品质量和患者安全。在此,我们采用了新一代测序技术,特别是RNA测序,并开发了一种方法来系统分析生产单克隆抗体的CHO细胞系mRNA的突变率。我们还仔细研究了延长培养时间以模拟生产过程中细胞扩增规模以及细胞培养中不同选择压力的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8fbc/4818804/7f2b7e53be03/BMRI2016-8356435.001.jpg

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