Multiplexed clonality verification of cell lines for protein biologic production.

ABSTRACT: During the development of cell lines for therapeutic protein production, a vector harboring a product transgene is integrated into the genome. To ensure production stability and consistent product quality, single-cell cloning is then performed. Since cells derived from the same parental clone have the same transgene integration locus, the identity of the integration site can also be used to verify the clonality of a production cell line. In this study, we present a high-throughput pipeline for clonality verification through integration site analysis. Sequence capture of genomic fragments that contain both vector and host cell genome sequences was used followed by next-generation sequencing to sequence the relevant vector-genome junctions. A Python algorithm was then developed for integrati

SUBMITTER: O'Brien SA 

PROVIDER: S-EPMC7803388 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature