title: Mutation Detection in an Antibody-Producing Chinese Hamster Ovary Cell Line by Targeted RNA Sequencing.
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.
authors.affiliation: Biologics & Vaccines, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Siyan
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Biology & Genetics Informatics, Merck Research Labs IT, Merck & Co, Boston, MA 02115, USA.
authors.forename: Jason D
authors.initials: JD
authors.lastname: Hughes
authors.affiliation: Discovery Pharmacogenomics, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Murgolo
authors.affiliation: Discovery Pharmacogenomics, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Diane
authors.initials: D
authors.lastname: Levitan
authors.affiliation: Biologics & Vaccines, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Janice
authors.initials: J
authors.lastname: Chen
authors.affiliation: Biologics & Vaccines, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Biologics & Vaccines, Merck Research Laboratories, Kenilworth, NJ 07033, USA.
authors.forename: Shuangping
authors.identifier: 0000-0002-7875-3569
authors.initials: S
authors.lastname: Shi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics*; D000917:Antibody Formation / Q000235:genetics*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009154:Mutation; D012313:RNA / Q000235:genetics*; D017423:Sequence Analysis, RNA
chemical_list: D000911:Antibodies, Monoclonal; D012313:RNA