title: High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system.
abstract: Therapeutic recombinant monoclonal antibodies (mAbs) are commonly produced by high-expressing, clonal, mammalian cells. Creation of these clones for manufacturing remains heavily reliant on stringent selection and gene amplification, which in turn can lead to genetic instability, variable expression, product heterogeneity and prolonged development timelines. Inclusion of cis-acting ubiquitous chromatin opening elements (UCOE™) in mammalian expression vectors has been shown to improve productivity and facilitate high-level gene expression irrespective of the chromosomal integration site without lengthy gene amplification protocols. In this study we have used high-throughput robotic clone selection in combination with UCOE™ containing expression vectors to develop a rapid, streamlined approach for early-stage cell line development and isolation of high-expressing clones for mAb production using Chinese hamster ovary (CHO) cells. Our results demonstrate that it is possible to go from transfection to stable clones in only 4 weeks, while achieving specific productivities exceeding 20 pg/cell/day. Furthermore, we have used this approach to quickly screen several process-crucial parameters including IgG subtype, enhancer-promoter combination and UCOE™ length. The use of UCOE™-containing vectors in combination with automated robotic selection provides a rapid method for the selection of stable, high-expressing clones.
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia. Electronic address: j.hou1@uq.edu.au.
authors: Jeff Jia Cheng
authors: JJ
authors: Hou
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors: Ben S
authors: BS
authors: Hughes
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors: Matthew
authors: M
authors: Smede
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors: Kar Man
authors: KM
authors: Leung
authors: Pharm Chemical Solution (PCS), EMD Millipore, 2 Gill Street, Woburn, MA 01801, USA.
authors: Kara
authors: K
authors: Levine
authors: Pharm Chemical Solution (PCS), EMD Millipore, 2 Gill Street, Woburn, MA 01801, USA.
authors: Susan
authors: S
authors: Rigby
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors: Peter P
authors: PP
authors: Gray
authors: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors: Trent P
authors: TP
authors: Munro
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001483:Base Sequence; D061249:Batch Cell Culture Techniques; D016466:CHO Cells; D002843:Chromatin / Q000378:metabolism*; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D005822:Genetic Vectors / Q000378:metabolism; D006168:Guinea Pigs; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D002843:Chromatin; D007074:Immunoglobulin G