High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system
•UCOE-containing cells show advantage in recombinant antibody expression.•Cell line generation involving the use of semi-solid cloning and ClonePix FL.•High throughput technology has a crucial role in clonal isolation.•Example of a clear, simple and streamlined path for biopharmaceutical development...
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Published in | New biotechnology Vol. 31; no. 3; pp. 214 - 220 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Netherlands
Elsevier B.V
25.05.2014
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Abstract | •UCOE-containing cells show advantage in recombinant antibody expression.•Cell line generation involving the use of semi-solid cloning and ClonePix FL.•High throughput technology has a crucial role in clonal isolation.•Example of a clear, simple and streamlined path for biopharmaceutical development.
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 20pg/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. |
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AbstractList | •UCOE-containing cells show advantage in recombinant antibody expression.•Cell line generation involving the use of semi-solid cloning and ClonePix FL.•High throughput technology has a crucial role in clonal isolation.•Example of a clear, simple and streamlined path for biopharmaceutical development.
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 20pg/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. 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. 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(TM)) 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(TM) 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(TM) length. The use of UCOE(TM)-containing vectors in combination with automated robotic selection provides a rapid method for the selection of stable, high-expressing clones. |
Author | Gray, Peter P. Rigby, Susan Hou, Jeff Jia Cheng Hughes, Ben S. Smede, Matthew Levine, Kara Munro, Trent P. Leung, Kar Man |
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Snippet | •UCOE-containing cells show advantage in recombinant antibody expression.•Cell line generation involving the use of semi-solid cloning and ClonePix FL.•High... Therapeutic recombinant monoclonal antibodies (mAbs) are commonly produced by high-expressing, clonal, mammalian cells. Creation of these clones for... |
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SubjectTerms | Animals Antibodies, Monoclonal - metabolism Base Sequence Batch Cell Culture Techniques CHO Cells Chromatin - metabolism Clone Cells Cricetinae Cricetulus Genetic Vectors - metabolism Guinea Pigs High-Throughput Screening Assays - methods Humans Immunoglobulin G - metabolism Promoter Regions, Genetic - genetics Transfection |
Title | High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system |
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