scholarly journals Tracing production instability in a clonally derived CHO cell line using single‐cell transcriptomics

2021 ◽  
Vol 118 (5) ◽  
pp. 2016-2030
Author(s):  
Ioanna Tzani ◽  
Nicholas Herrmann ◽  
Sara Carillo ◽  
Cathy A. Spargo ◽  
Ryan Hagan ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
Cho Cell ◽  
Cho Cell Line

scholarly journals Tracing production instability in a clonally-derived CHO cell line using single cell transcriptomics

2020 ◽  
Author(s):  
Ioanna Tzani ◽  
Nicolas Herrmann ◽  
Sara Carillo ◽  
Cathy A. Spargo ◽  
Ryan Hagan ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Monoclonal Antibody ◽  
Single Cell ◽  
Cell Line ◽  
Light Chain ◽  
Chain Gene ◽  
Cho Cell ◽  
Light Chain Gene ◽  
Cho Cell Line

AbstractA variety of mechanisms including transcriptional silencing, gene copy loss and increased susceptibility to cellular stress have been associated with a sudden or gradual loss of monoclonal antibody (mAb) production in Chinese hamster ovary (CHO) cell lines. In this study, we utilised single cell RNASeq (scRNASeq) to study a clonally-derived CHO cell line that underwent production instability leading to a dramatic reduction of the levels of mAb produced. From the scRNASeq data we identified sub clusters associated with variations in the mAb transgenes and observed that heavy chain gene expression was significantly lower than that of the light chain across the population. Using trajectory inference, the evolution of the cell line was reconstructed and was found to correlate with a reduction in heavy and light chain gene expression. Genes encoding for proteins involved in the response to oxidative stress and apoptosis were found to increase in expression as cells progressed along the trajectory. Future studies of CHO cell lines using this technology have the potential to dramatically enhance our understanding of the characteristics underpinning efficient manufacturing performance as well as product quality.HighlightsA clonally-derived CHO cell line in our laboratory had undergone production instability – in that the amount of intact monoclonal antibody had reduced dramatically to levels at which reliable quantitation was no longer possible. We were, however, able to detect mAb heavy and light chain protein, as well as dimerised light chain species in the cell culture media.Single cell RNASeq was utilised to capture > 3,800 gene expression profiles from the cell line at 72hrs post seeding.Analyses of the scRNASeq data uncovered transcriptional heterogeneity and revealed the presence of multiple intra cell line clusters. The heavy chain transcript was detected at a significantly lower level in comparison light chain transcripts. Light chain gene expression was not only more abundant, but also expressed more uniformly across the cell population.Using unsupervised trajectory analysis, the emergence of heterogeneity in the cell population was traced from those cells most similar to the original isolated clone to those where transcription of the mAb heavy and light chain was undetectable.Subsequent analysis of CHO cell gene expression patterns revealed a correlation between the progression of cells along the trajectory and the upregulation of genes involved in the cellular response to oxidative stress.

scholarly journals A framework to quantify karyotype variation associated with CHO cell line instability at a single-cell level

2017 ◽  
Vol 114 (5) ◽  
pp. 1045-1053 ◽  
Author(s):  
Jong Youn Baik ◽  
Kelvin H. Lee
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
Single Cell Level ◽  
Cho Cell ◽  
Cell Level ◽  
Cho Cell Line

Leveraging a CHO cell line toolkit to accelerate biotherapeutics into the clinic

2017 ◽  
Vol 33 (6) ◽  
pp. 1468-1475 ◽  
Author(s):  
Chapman Wright ◽  
Christina Alves ◽  
Rashmi Kshirsagar ◽  
John Pieracci ◽  
Scott Estes
Keyword(s):  
Cell Line ◽  
Cho Cell ◽  
Cho Cell Line

scholarly journals High performance CHO cell line development platform for enhanced production of recombinant proteins including difficult-to-express proteins

2013 ◽  
Vol 7 (S6) ◽  
Author(s):  
Pierre-Alain Girod ◽  
Valérie Le Fourn ◽  
David Calabrese ◽  
Alexandre Regamey ◽  
Deborah Ley ◽  
...  
Keyword(s):  
Cell Line ◽  
Recombinant Proteins ◽  
High Performance ◽  
Cho Cell ◽  
Cell Line Development ◽  
Development Platform ◽  
Enhanced Production ◽  
Cho Cell Line

O-109 A first dose-response trial investigating the effects of adding choriogonadotropin beta to follitropin delta in women undergoing ovarian stimulation in a long GnRH agonist protocol

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Fernandez Sanchez ◽  
H Višnová ◽  
C Blockeel ◽  
A Pinborg ◽  
Y Khalaf ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Cell Line ◽  
Gnrh Agonist ◽  
Ovarian Stimulation ◽  
Randomised Trial ◽  
Chinese Hamster ◽  
Ongoing Pregnancy ◽  
Cho Cell ◽  
Ongoing Pregnancy Rate ◽  
Cho Cell Line

Abstract Study question Does addition of choriogonadotropin beta (CG beta) to follitropin delta increase the number of good-quality blastocysts following ovarian stimulation in a long GnRH agonist protocol? Summary answer At the doses investigated, the addition of CG beta reduced the number of intermediate follicles and decreased the number of oocytes and blastocysts. What is known already CG beta is a new recombinant hCG (rhCG) molecule expressed by a human cell line (PER.C6â) with a different glycosylation profile compared to urinary hCG or rhCG derived from a Chinese Hamster Ovary (CHO) cell-line. In the first-in-human trial, the CG beta pharmacokinetics were similar between men and women. In women, the area under the curve (AUC) and the peak serum concentration (Cmax) increased dose proportionally following single and multiple daily doses. In men, a single dose of CG beta provided higher exposure with a longer half-life and proportionately higher testosterone production than rhCG derived from a CHO cell line. Study design, size, duration Placebo-controlled, double-blind, randomised trial (RAINBOW) to explore the efficacy and safety of CG beta as add-on treatment to follitropin delta in women undergoing COS in a long GnRH agonist protocol. The primary endpoint was the number of good-quality blastocysts (grade 3 BB or higher, Gardner and Schoolcraft, 1999). Subjects were randomised to receive either placebo or 1, 2, 4, 8, or 12 µg CG beta added to the daily individualised follitropin delta dose during COS. Participants/materials, setting, methods In total 619 women (30-42 years) with AMH levels between 5 and 35 pmol/L were randomized in equal proportions to the six treatment groups. All subjects were treated with an individualised dose of follitropin delta determined based on AMH (Elecsys AMH Plus Immunoassay) and body weight. Triggering was performed when 3 follicles were ≥17 mm but no more than 25 follicles ≥12 mm were reached Main results and the role of chance The incidence of cycle cancellation (range 0%-2.9%), total follitropin delta dose (mean 112 µg) and duration of stimulation (mean 10 days) were similar across the groups. A reduced number of intermediate follicles (12 to 17 mm) and fewer oocytes (mean range 9.7 to 11.2) were observed for all doses of CG beta compared to the follitropin delta only group (mean 12.5). The mean number of goodquality blastocysts was 3.3 in the follitropin delta group and ranged between 2.1 and 3.0 across the CG beta groups. The incidence of transfer cancellation was higher in the 4, 8 and 12 µg group, mostly as no blastocyst was available for transfer. In the group receiving only follitropin delta, the ongoing pregnancy rate (10-11 weeks after transfer) was high i.e. 43% per started cycle vs 28-39% in CG beta groups and 49% per transfer vs 38-50% in the CG beta groups. In line with the number of collected oocytes, the OHSS incidence was overall lower following follitropin delta with CG beta than following follitropin delta only treatment. Regardless of the dose, CG beta was safe and well-tolerated with low risk of immunogenicity. Limitations, reasons for caution The effect of the unique glycosylation of CG beta and the associated potency implications in women were not known prior to this trial. Further studies will be needed to evaluate potentially lower doses of CG beta for this and/or different indications. Wider implications of the findings The high ongoing pregnancy rate in the follitropin delta group supports the use of individualised follitropin delta dosing in a long GnRH agonist protocol. The differential potency of CG beta may have impaired the growth of intermediate follicles with the investigated doses without affecting the ongoing pregnancy rates per transfer. Trial registration number NCT03564509

Perfusion Cultivation of Genetically-Engineered CHO Cell Line Producing Prourokinase with Home-Made SH-2 Microcarrier and Low-Serum Medium

1997 ◽  
pp. 255-260
Author(s):  
Chengzu Xiao ◽  
Zicai Huang ◽  
Zhaolie Chen ◽  
Zhixia Guo ◽  
Lihua Gao ◽  
...  
Keyword(s):  
Cell Line ◽  
Genetically Engineered ◽  
Cho Cell ◽  
Cho Cell Line ◽  
Perfusion Cultivation ◽  
Low Serum

Process intensification for the production of rituximab by an inducible CHO cell line

2019 ◽  
Vol 42 (5) ◽  
pp. 711-725 ◽  
Author(s):  
Kahina Mellahi ◽  
Denis Brochu ◽  
Michel Gilbert ◽  
Michel Perrier ◽  
Sven Ansorge ◽  
...  
Keyword(s):  
Cell Line ◽  
Process Intensification ◽  
Cho Cell ◽  
Cho Cell Line

Bioprocess development of a stable FUT8−/−-CHO cell line to produce defucosylated anti-HER2 antibody

2019 ◽  
Vol 42 (8) ◽  
pp. 1263-1271 ◽  
Author(s):  
Yuan Yuan ◽  
Huifang Zong ◽  
Jingyi Bai ◽  
Lei Han ◽  
Lei Wang ◽  
...  
Keyword(s):  
Cell Line ◽  
Bioprocess Development ◽  
Cho Cell ◽  
Cho Cell Line

Using chemical chaperones to increase recombinant human erythropoietin secretion in CHO cell line

2019 ◽  
Vol 49 (6) ◽  
pp. 535-544 ◽  
Author(s):  
Mehri Mortazavi ◽  
Mohammad Ali Shokrgozar ◽  
Soroush Sardari ◽  
Kayhan Azadmanesh ◽  
Reza Mahdian ◽  
...  
Keyword(s):  
Cell Line ◽  
Recombinant Human Erythropoietin ◽  
Cho Cell ◽  
Chemical Chaperones ◽  
Human Erythropoietin ◽  
Cho Cell Line
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