scholarly journals Study of a recombinant CHO cell line producing a monoclonal antibody by ATF or TFF external filter perfusion in a WAVE Bioreactor™

2011 ◽  
Vol 5 (Suppl 8) ◽  
pp. P105 ◽  
Author(s):  
Marie-Françoise Clincke ◽  
Carin Mölleryd ◽  
Ye Zhang ◽  
Eva Lindskog ◽  
Kieron Walsh ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Line ◽  
Cho Cell ◽  
Wave Bioreactor ◽  
Cho Cell Line ◽  
Recombinant Cho Cell Line

Evaluation of the Virus Elimination and Inactivation at Different Stages of the Model Biotechnological Process for the Production of a Drug Based on the Monoclonal Antibody Fab-Fragment

2021 ◽  
Vol 37 (1) ◽  
pp. 69-80
Author(s):  
S.G. Yurkov ◽  
S.P Zhivoderov ◽  
A.Y. Koltsov ◽  
R.A. Khamitov ◽  
N.V. Stratonova ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Line ◽  
Encephalomyocarditis Virus ◽  
Virus Infectivity ◽  
Cho Cell ◽  
Fab Fragment ◽  
Virus Elimination ◽  
Virus Removal ◽  
Cho Cell Line ◽  
Virus Safety

Virus safety insuring is one of the most serious problems in the development of biotechnological drugs produced using animal cell lines. Quantitative assessment of virus removal and inactivation is an integral approach to show the reliability of the target compound production. In this work, the model experiments have been carried out on the purification of the monoclonal antibody recombinant Fab-fragment from viruses of various origins and properties: xenotropic murine leukemia virus (X-MuLV), pseudorabies virus (PRV), Reo-3 virus and encephalomyocarditis virus (EMCV). It was shown that two methods, acidification to pH 3.2 and nanofiltration, made it possible to reduce the virus infectivity to levels undetectable in cell cultures (according to TCID50). The developed multistage purification process of the target protein provided an overall decrease in viral clearance to the following values: X-MuLV≥10.17lg, PRV≥13.98lg, Reo-3≥8.09lg and EMCV≥4.98lg. These results confirm that the developed technology ensures the virus safety during the production of a monoclonal antibody recombinant Fab-fragment by CHO cell line. These results confirm virus safety of production technology of recombinant monoclonal antibody Fab-fragment produced in CHO cell line. virus safety, virus elimination, virus inactivation, nanofiltration, Fab-fragment, monoclonal antibody, CHO cell line

High-Level scu-PA Production by Butyrate-Treated Serum-Free Culture of Recombinant CHO Cell Line

2004 ◽  
Vol 20 (6) ◽  
pp. 1788-1796 ◽  
Author(s):  
J.-S. Kim ◽  
B.-C. Ahn ◽  
B.-P. Lim ◽  
Y.D. Choi ◽  
E.-C. Jo
Keyword(s):  
Cell Line ◽  
Cho Cell ◽  
Serum Free ◽  
Cho Cell Line ◽  
High Level ◽  
Recombinant 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.

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