High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system

Hou, Jeff Jia Cheng, Hughes, Ben S., Smede, Matthew, Leung, Kar Man, Levine, Kara, Rigby, Susan, Gray, Peter P. and Munro, Trent P. (2014) High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system. New Biotechnology, 31 3: 214-220. doi:10.1016/j.nbt.2014.02.002


Author Hou, Jeff Jia Cheng
Hughes, Ben S.
Smede, Matthew
Leung, Kar Man
Levine, Kara
Rigby, Susan
Gray, Peter P.
Munro, Trent P.
Title High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system
Journal name New Biotechnology   Check publisher's open access policy
ISSN 1871-6784
1876-4347
Publication date 2014-05
Year available 2014
Sub-type Article (original research)
DOI 10.1016/j.nbt.2014.02.002
Open Access Status
Volume 31
Issue 3
Start page 214
End page 220
Total pages 7
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2015
Language eng
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.
Keyword Hamster ovary cells
Mammalian cells
Gene amplification
Selective pressure
Protein production
Cho cells
Stability
Line
Therapeutics
Vectors
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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