title: A high-throughput automated platform for the development of manufacturing cell lines for protein therapeutics.
abstract: The fast-growing biopharmaceutical industry demands speedy development of highly efficient and reliable production systems to meet the increasing requirement for drug supplies. The generation of production cell lines has traditionally involved manual operations that are labor-intensive, low-throughput and vulnerable to human errors. We report here an integrated high-throughput and automated platform for development of manufacturing cell lines for the production of protein therapeutics. The combination of BD FACS Aria Cell Sorter, CloneSelect Imager and TECAN Freedom EVO liquid handling system has enabled a high-throughput and more efficient cell line development process. In this operation, production host cells are first transfected with an expression vector carrying the gene of interest (1), followed by the treatment with a selection agent. The stably-transfected cells are then stained with fluorescence-labeled anti-human IgG antibody, and are subsequently subject to flow cytometry analysis (2-4). Highly productive cells are selected based on fluorescence intensity and are isolated by single-cell sorting on a BD FACSAria. Colony formation from single-cell stage was detected microscopically and a series of time-laps digital images are taken by CloneSelect Imager for the documentation of cell line history. After single clones have formed, these clones were screened for productivity by ELISA performed on a TECAN Freedom EVO liquid handling system. Approximately 2,000 - 10,000 clones can be screened per operation cycle with the current system setup. This integrated approach has been used to generate high producing Chinese hamster ovary (CHO) cell lines for the production of therapeutic monoclonal antibody (mAb) as well as their fusion proteins. With the aid of different types of detecting probes, the method can be used for developing other protein therapeutics or be applied to other production host systems. Comparing to the traditional manual procedure, this automated platform demonstrated advantages of significantly increased capacity, ensured clonality, traceability in cell line history with electronic documentation and much reduced opportunity in operator error.
authors.affiliation: Merck Research Laboratory, Merck & Co., Inc. shuangping.shi@merck.com
authors.forename: Shuangping
authors.initials: S
authors.lastname: Shi
authors.forename: Russ G G
authors.initials: RG
authors.lastname: Condon
authors.forename: Liang
authors.initials: L
authors.lastname: Deng
authors.forename: Jason
authors.initials: J
authors.lastname: Saunders
authors.forename: Finn
authors.initials: F
authors.lastname: Hung
authors.forename: Yung-Shyeng
authors.initials: YS
authors.lastname: Tsao
authors.forename: Zhong
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001702:Biopharmaceutics / Q000379:methods*; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003584:Cytological Techniques / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005434:Flow Cytometry; D057166:High-Throughput Screening Assays / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal