Identification and control of novel growth inhibitors in fed-batch cultures of Chinese hamster ovary cells

2017 ◽  
Vol 114 (8) ◽  
pp. 1779-1790 ◽  
Author(s):  
Bhanu Chandra Mulukutla ◽  
Jaitashree Kale ◽  
Taylor Kalomeris ◽  
Michaela Jacobs ◽  
Gregory W. Hiller
Keyword(s):  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Growth Inhibitors ◽  
Fed Batch ◽  
Ovary Cells ◽  
Batch Cultures ◽  
And Control

Metabolic engineering of Chinese hamster ovary cells towards reduced biosynthesis and accumulation of novel growth inhibitors in fed-batch cultures

2019 ◽  
Vol 54 ◽  
pp. 54-68 ◽  
Author(s):  
Bhanu Chandra Mulukutla ◽  
Jeffrey Mitchell ◽  
Pauline Geoffroy ◽  
Cameron Harrington ◽  
Manisha Krishnan ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Growth Inhibitors ◽  
Fed Batch ◽  
Ovary Cells ◽  
Batch Cultures

scholarly journals Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses

iScience ◽  
2019 ◽  
Vol 12 ◽  
pp. 102-120 ◽  
Author(s):  
Madhuresh Sumit ◽  
Sepideh Dolatshahi ◽  
An-Hsiang Adam Chu ◽  
Kaffa Cote ◽  
John J. Scarcelli ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Time Course ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Fed Batch ◽  
Ovary Cells ◽  
Batch Cultures

Dissecting the Dynamics of N-Linked Glycosylation in Fed-Batch Cultures of Chinese Hamster Ovary Cells Through Time Course Omics Analyses

2018 ◽  
Author(s):  
Madhuresh Sumit ◽  
Sepideh Dolatshahi ◽  
An-Hsiang Adam Chu ◽  
Kaffa Cote ◽  
John J. Scarcelli ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Time Course ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Fed Batch ◽  
Ovary Cells ◽  
Batch Cultures

scholarly journals Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production

2020 ◽  
Vol 245 (8) ◽  
pp. 690-702
Author(s):  
Xiaonan Ma ◽  
Ling Zhang ◽  
Luming Zhang ◽  
Chenglong Wang ◽  
Xiaorui Guo ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Chinese Hamster Ovary ◽  
Reference Genes ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Expression Stability ◽  
Fed Batch ◽  
Ovary Cells ◽  
Fc Fusion Protein

Chinese hamster ovary cells are the predominant cell lines used for bio-therapeutic production. Real-time quantitative PCR (RT-qPCR) and transcriptomics are powerful tools to understand and optimize the Chinese hamster ovary cells for higher productivity or better control of product qualities. Reliable reference genes, which were proved to be experiment-specific, are critical yardsticks. In this study, we compared expression stability of 20 candidate reference genes at mRNA level, including commonly used housekeeping genes, previous literature reported genes in Chinese hamster ovary cells producing an intact antibody, and new candidates suggested by our RNA-seq transcriptomic database, in RT-qPCR reactions in Fc-fusion protein-producing Chinese hamster ovary cells with various productivity during long-term cultivation and fed-batch cultures at 26 different conditions. geNorm, NormFinder, BestKeeper, and ΔCt programs and methods were utilized to analyze the gene expression stability and gave an overall ranking. Akr1a1, Gpx1, and Aprt in long-term cultivation and Akr1a1, Rps16 in fed-batch culture, which have not been reported previously, exhibited the highest stability of gene expression, while Pabpn1, Hirip3, and Actb in both sets of experiments together with Atp5f1 in long-term passage process showed the weakest stability. The results were then validated using GLP1-Fc (Glucagon-like peptide-1 Fc fusion protein) gene as the target with determined expression level which were doubly confirmed by both absolute RT-qPCR and confocal microscopy. These new references should be considered for the investigations on Chinese hamster ovary cells in related research. Impact statement In order to reveal potential genotype-phenotype relationship, RT-qPCR reactions are frequently applied which require validated and reliable reference genes. With the investigation on long-term passage and fed-batch cultivation of CHO cells producing an Fc-fusion protein, four new reference genes–Akr1a1, Gpx1, Aprt, and Rps16, were identified from 20 candidates with the aid of geNorm, NormFinder, BestKeeper, and ΔCt programs and methods. This article provided more verified options in reference gene selection in related research on CHO cells.

scholarly journals Enhanced recombinant factor VII expression in Chinese hamster ovary cells by optimizing signal peptides and fed-batch medium

Bioengineered ◽  
2016 ◽  
Vol 7 (3) ◽  
pp. 189-197 ◽  
Author(s):  
Lin Peng ◽  
Xiao Yu ◽  
Chengyuan Li ◽  
Yanfei Cai ◽  
Yun Chen ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Factor Vii ◽  
Signal Peptides ◽  
Fed Batch ◽  
Ovary Cells
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