Cell Xpress™ Technology Facilitates High-Producing Chinese Hamster Ovary Cell Line Generation Using Glutamine Synthetase Gene Expression System

Richardson, Genova A.; Lin, Nan; Lacy, Kimberly A.; Davis, Lynn; Gray, Misa I.; Cresswell, Jennifer; Gerber, Mark A.; Caple, Matthew V.; Kayser, Kevin J.

doi:10.1007/978-90-481-3419-9_9

Cited by 4 publications

(2 citation statements)

References 2 publications

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“…Several methods such as Cell Xpress technology, 29 transcriptome-based mRNA screening, and fluorescence-activated cell sorting (FACS)-based selection systems have been used to increase the identification of highproducing clones in a less labor-intensive way. [31][32][33][34] Another approach is to increase the selection stringency to improve the percentage of high-producing clones.…”

Section: Discussionmentioning

confidence: 99%

Attenuated glutamine synthetase as a selection marker in CHO cells to efficiently isolate highly productive stable cells for the production of antibodies and other biologics

Lin

Chan

Kieß

et al. 2019

mAbs

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Chinese hamster ovary (CHO) cells are the biopharmaceutical industry's primary means of manufacturing therapeutic proteins, including monoclonal antibodies. The major challenge in cell line development for the production of recombinant biopharmaceuticals lies in generating and isolating rare high-producing stable clones, amongst thousands of low-producing or unstable clones, in a short period of time. One approach to accomplish this is to use the glutamine synthetase (GS) selection system, together with the GS inhibitor, methionine sulfoximine (MSX). However, MSX can only increase protein productivity to a limited extent. Often productivity will drop when MSX is removed from the system. We evaluated a congenital GS mutation, R324C, which causes glutamine deficiency in human as an attenuated selection marker for CHO cell line generation. We also created a panel of GS mutants with diminished GS activity. Our results demonstrated that using attenuated GS mutants as selection markers significantly increased antibody production of stably transfected pools. Furthermore, these stably transfected pools sustained high productivity levels for an extended period of time, whereas cells transfected with wild-type GS lost considerable protein productivity over time, particularly after MSX was removed. In summary, the use of attenuated GS as a selection marker in CHO cell line development bypasses the need for MSX, and generates stable clones with significantly higher antibody productivity.

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Section: Discussionmentioning

confidence: 99%

Attenuated glutamine synthetase as a selection marker in CHO cells to efficiently isolate highly productive stable cells for the production of antibodies and other biologics

Lin

Chan

Kieß

et al. 2019

mAbs

View full text Add to dashboard Cite

show abstract

“…Furthermore, as the entire process requires less than 30 s to screen a single well, large numbers of cell clones can be screened within a short time to identify high producers. It has also been reported that the analysis result from Cell Xpress TM technology shares a good correlation (R 2 = 0.84) with the peak IgG volumetric productivities in shake flask growth and expression experiments [116]. In general, Cell Xpress TM technology has routinely picked cell lines with specific antibody secretion rates of >50pg/cell per day [114].…”

Section: Clone Screening Technologiesmentioning

confidence: 99%

Advances in Mammalian Cell Line Development Technologies for Recombinant Protein Production

2013

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From 2006 to 2011, an average of 15 novel recombinant protein therapeutics have been approved by US Food and Drug Administration (FDA) annually. In addition, the expiration of blockbuster biologics has also spurred the emergence of biosimilars. The increasing numbers of innovator biologic products and biosimilars have thus fuelled the demand of production cell lines with high productivity. Currently, mammalian cell line development technologies used by most biopharmaceutical companies are based on either the methotrexate (MTX) amplification technology or the glutamine synthetase (GS) system. With both systems, the cell clones obtained are highly heterogeneous, as a result of random genome integration by the gene of interest and the gene amplification process. Consequently, large numbers of cell clones have to be screened to identify rare stable high producer cell clones. As such, the cell line development process typically requires 6 to 12 months and is a time, capital and labour intensive process. This article reviews established advances in protein expression and clone screening which are the core technologies in mammalian cell line development. Advancements in these component technologies are vital to improve the speed and efficiency of generating robust and highly productive cell line for large scale production of protein therapeutics.

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Recombinant expression systems

2015

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Cell Xpress™ Technology Facilitates High-Producing Chinese Hamster Ovary Cell Line Generation Using Glutamine Synthetase Gene Expression System

Cited by 4 publications

References 2 publications

Attenuated glutamine synthetase as a selection marker in CHO cells to efficiently isolate highly productive stable cells for the production of antibodies and other biologics

Attenuated glutamine synthetase as a selection marker in CHO cells to efficiently isolate highly productive stable cells for the production of antibodies and other biologics

Advances in Mammalian Cell Line Development Technologies for Recombinant Protein Production

Recombinant expression systems

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