IntroductionChinese hamster ovary (CHO) cells continue to be the cell factory of choice for large‐scale biopharmaceutical production. To produce a therapeutic protein with the required quality properties according to FDA and EMA regulations, a stable clonal cell line has to be generated, enabling long‐term and reproducible protein expression. Therefore, the classical cell line generation procedure consists of random integration of the transgene into the genome of host cells, followed by selection of transgene containing cells using antibiotics or other transgene selection strategies like methionine sulfoximine (MSX). Subsequently, single‐cell cloning and finally, selection of high‐producer clones from the bulk of low‐ and non‐producers are performed. Selection of high‐producer clones remains one of the major bottlenecks in cell line development (CLD), taking up 4 to 8 months and consuming valuable financial and human resources. Random transgene integration, different transgene copy numbers, and the selection pressure encountered during CLD all contribute to genetically heterogeneous cell populations that require efficient screening processes. Several strategies have been developed in recent years to improve the ease, throughput and performance of the selection processes, carefully reviewed by Priola et al. These strategies are to a certain degree used in academic labs, but even more so in
Biotechnology Journal – Wiley
Published: Jan 1, 2018
Keywords: ; ; ; ; ;
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