Changes in Chromosome Counts and Patterns in CHO Cell Lines upon Generation of Recombinant Cell Lines and Subcloning

2018 ◽  
Vol 13 (3) ◽  
pp. 1700495 ◽  
Author(s):  
Sabine Vcelar ◽  
Michael Melcher ◽  
Norbert Auer ◽  
Astrid Hrdina ◽  
Anja Puklowski ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cho Cell ◽  
Chromosome Counts ◽  
Recombinant Cell

scholarly journals Recombinant γY278H Fibrinogen Showed Normal Secretion from CHO Cells, but a Corresponding Heterozygous Patient Showed Hypofibrinogenemia

2021 ◽  
Vol 22 (10) ◽  
pp. 5218
Author(s):  
Tomu Kamijo ◽  
Takahiro Kaido ◽  
Masahiro Yoda ◽  
Shinpei Arai ◽  
Kazuyoshi Yamauchi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cho Cells ◽  
Fibrinogen Level ◽  
Culture Media ◽  
Chinese Hamster ◽  
Plasma Fibrinogen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cho Cell ◽  
Accelerated Degradation ◽  
Heterozygous Patient

We identified a novel heterozygous hypofibrinogenemia, γY278H (Hiroshima). To demonstrate the cause of reduced plasma fibrinogen levels (functional level: 1.12 g/L and antigenic level: 1.16 g/L), we established γY278H fibrinogen-producing Chinese hamster ovary (CHO) cells. An enzyme-linked immunosorbent assay demonstrated that synthesis of γY278H fibrinogen inside CHO cells and secretion into the culture media were not reduced. Then, we established an additional five variant fibrinogen-producing CHO cell lines (γL276P, γT277P, γT277R, γA279D, and γY280C) and conducted further investigations. We have already established 33 γ-module variant fibrinogen-producing CHO cell lines, including 6 cell lines in this study, but only the γY278H and γT277R cell lines showed disagreement, namely, recombinant fibrinogen production was not reduced but the patients’ plasma fibrinogen level was reduced. Finally, we performed fibrinogen degradation assays and demonstrated that the γY278H and γT277R fibrinogens were easily cleaved by plasmin whereas their polymerization in the presence of Ca2+ and “D:D” interaction was normal. In conclusion, our investigation suggested that patient γY278H showed hypofibrinogenemia because γY278H fibrinogen was secreted normally from the patient’s hepatocytes but then underwent accelerated degradation by plasmin in the circulation.

scholarly journals Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Laura Bryan ◽  
Michael Henry ◽  
Ronan M. Kelly ◽  
Christopher C. Frye ◽  
Matthew D. Osborne ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Density ◽  
Cell Lines ◽  
Viable Cell ◽  
Viable Cell Density ◽  
High Peak ◽  
Proteomic Profiling ◽  
Cho Cell ◽  
Extended Culture ◽  
Key Pathways

Abstract Background The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods. In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. Results We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. Conclusion This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production.

scholarly journals A strategy to obtain recombinant cell lines with high expression levels. Lentiviral vector-mediated transgenesis

2011 ◽  
Vol 5 (S8) ◽  
Author(s):  
Claudio Prieto ◽  
Diego Fontana ◽  
Marina Etcheverrigaray ◽  
Ricardo Kratje
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
High Expression ◽  
Expression Levels ◽  
Recombinant Cell

Interferon-?-(IFN) producing CHO cell lines are resistant to the antiproliferative activity of IFN: A correlation with gene expression

1988 ◽  
Vol 38 (4) ◽  
pp. 269-278 ◽  
Author(s):  
M. van Heuvel ◽  
M. Govaert-Siemerink ◽  
I. J. Bosveld ◽  
E. G. Zwarthoff ◽  
J. Trapman
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Antiproliferative Activity ◽  
Cho Cell

P IX.17 Homologous recombination in CHO cell lines defective in DNA damage processing

1997 ◽  
Vol 379 (1) ◽  
pp. S65
Author(s):  
M. Kruszewski ◽  
H. Kruszewska ◽  
H. Inaba ◽  
P. Jeggo ◽  
I. Szumiel
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Cell Lines ◽  
Cho Cell

scholarly journals Generation of Flp-intm-ready DG44 and Lec 3.2.8.1 CHO cell lines for quick and easy constitutive protein expression

BioTechniques ◽  
2018 ◽  
Vol 65 (1) ◽  
pp. 41-46 ◽  
Author(s):  
DC Soler ◽  
AE Young ◽  
A Vahedi-Faridi ◽  
TS McCormick
Keyword(s):  
Protein Expression ◽  
Cell Lines ◽  
Cho Cell

Strategies for selecting Recombinant CHO cell lines for cGMP manufacturing: Realizing the potential in bioreactors

2010 ◽  
Vol 26 (5) ◽  
pp. 1446-1454 ◽  
Author(s):  
Alison J. Porter ◽  
Alan J. Dickson ◽  
Andrew J. Racher
Keyword(s):  
Cell Lines ◽  
Cho Cell
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