Limitations to the Development of Humanized Antibody Producing Chinese Hamster Ovary Cells Using Glutamine Synthetase-Mediated Gene Amplification

2006 ◽  
Vol 22 (3) ◽  
pp. 770-780 ◽  
Author(s):  
S.C. Jun ◽  
M.S. Kim ◽  
H.J. Hong ◽  
G.M. Lee
Keyword(s):  
Glutamine Synthetase ◽  
Gene Amplification ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Humanized Antibody

Effect of culture temperature on TNFR-Fc productivity in recombinant glutamine synthetase-chinese hamster ovary cells

2010 ◽  
Vol 32 (9) ◽  
pp. 1239-1244 ◽  
Author(s):  
Li Fan ◽  
Liang Zhao ◽  
Zhaoyang Ye ◽  
Yating Sun ◽  
Tianci Kou ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Culture Temperature ◽  
Ovary Cells

Amplification and expression of human alpha-globin genes in Chinese hamster ovary cells

1984 ◽  
Vol 4 (8) ◽  
pp. 1469-1475
Author(s):  
Y F Lau ◽  
C C Lin ◽  
Y W Kan
Keyword(s):  
Gene Expression ◽  
Gene Amplification ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Globin Gene ◽  
Chinese Hamster ◽  
Globin Genes ◽  
Ovary Cells ◽  
Alpha Globin ◽  
Globin Gene Expression

We studied the effects of gene amplification on human globin gene expression in Chinese hamster ovary cells. The normal human alpha-globin gene (N alpha 2) and a hybrid gene (M alpha G) containing the 5' promoter-regulator region of the mouse metallothionein gene and the human structural alpha 2-globin gene were linked to a modular SV2-cDNA dihydrofolate reductase (DHFR) gene. The recombinant DNA molecules were introduced into Chinese hamster ovary cells by calcium phosphate precipitation. After initial selection to retain the DHFR and linked sequences, the cells were cultured in increasing concentrations of methotrexate up to 0.2 mM. Southern blot analysis of total cellular DNA showed an approximately 500- to 1,000-fold increase in the number of copies of DHFR and human alpha-globin genes. The transcription of the alpha-globin and DHFR genes increased as their copy number within the cells increased. The transcription of the amplified hybrid M alpha G gene was also inducible with cadmium treatments. Both mature mRNA and "read-through" transcripts were observed. DHFR constituted approximately 10% of pulse-labeled cellular proteins in these cells, but no human alpha-globin was detected. In vitro translation of polyadenylated RNA from these cells showed that alpha-globin mRNA transcribed from the amplified alpha-globin genes was functional and directed alpha-globin chain synthesis. In situ hybridization of 3H-labeled alpha-globin and DHFR DNA probes in chromosome preparations from the two cell lines indicated that both genes were coamplified in the same chromosomal locations in each cell type. These results indicate that gene amplification enhances human globin gene expression in cultured Chinese hamster ovary cells.

Effects of palindrome structure on dihydrofolate reductase gene amplification in Chinese hamster ovary cells

2009 ◽  
Vol 108 ◽  
pp. S9
Author(s):  
Joon Young Park ◽  
Yasuhiro Takagi ◽  
Miyuki Yamatani ◽  
Kohsuke Honda ◽  
Takeshi Omasa ◽  
...  
Keyword(s):  
Gene Amplification ◽  
Dihydrofolate Reductase ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Reductase Gene

Improved gene amplification by cell-cycle engineering combined with the Cre-loxP system in Chinese hamster ovary cells

2015 ◽  
Vol 120 (6) ◽  
pp. 701-708 ◽  
Author(s):  
Rima Matsuyama ◽  
Tomomi Tsutsui ◽  
Kyoung Ho Lee ◽  
Masayoshi Onitsuka ◽  
Takeshi Omasa
Keyword(s):  
Cell Cycle ◽  
Gene Amplification ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells
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