scholarly journals Mutation Detection in an Antibody-Producing Chinese Hamster Ovary Cell Line by Targeted RNA Sequencing

2016 ◽  
Vol 2016 ◽  
pp. 1-8
Author(s):  
Siyan Zhang ◽  
Jason D. Hughes ◽  
Nicholas Murgolo ◽  
Diane Levitan ◽  
Janice Chen ◽  
...  

Chinese hamster ovary (CHO) cells have been used widely in the pharmaceutical industry for production of biological therapeutics including monoclonal antibodies (mAb). The integrity of the gene of interest and the accuracy of the relay of genetic information impact product quality and patient safety. Here we employed next-generation sequencing, particularly RNA-seq, and developed a method to systematically analyze the mutation rate of the mRNA of CHO cell lines producing a mAb. The effect of an extended culturing period to mimic the scale of cell expansion in a manufacturing process and varying selection pressure in the cell culture were also closely examined.

2011 ◽  
Vol 156 (3) ◽  
pp. 227-235 ◽  
Author(s):  
Jennifer Becker ◽  
Matthias Hackl ◽  
Oliver Rupp ◽  
Tobias Jakobi ◽  
Jessica Schneider ◽  
...  

1989 ◽  
Vol 9 (4) ◽  
pp. 1754-1758
Author(s):  
T M Underhill ◽  
W F Flintoff

A methotrexate-resistant Chinese hamster ovary cell line deficient in methotrexate uptake has been complemented to methotrexate sensitivity by transfection with DNA isolated from either wild-type Chinese hamster ovary or human G2 cells. Primary and secondary transfectants regained the ability to take up methotrexate in a manner similar to that of wild-type cells, and in the case of those transfected with human DNA, to contain human-specific DNA sequences. The complementation by DNA-mediated gene transfer of this methotrexate-resistant phenotype provides a basis for the cloning of a gene involved in methotrexate uptake.


1981 ◽  
Vol 1 (12) ◽  
pp. 1069-1076 ◽  
Author(s):  
R J Kaufman ◽  
R T Schimke

During stepwise increases in the methotrexate concentration in culture medium, we selected Chinese hamster ovary cells that contained elevated dihydrofolate reductase levels which were proportional to the number of dihydrofolate reductase gene copies (i.e., gene amplification). We studied the dihydrofolate reductase levels in individual cells that underwent the initial steps of methotrexate resistance by using the fluorescence-activated cell sorter technique. Such cells constituted a heterogeneous population with differing dihydrofolate reductase levels, and they characteristically lost the elevated enzyme levels when they were grown in the absence of methotrexate. The progeny of individual cells with high enzyme levels behaved differently and could lose all or variable numbers of the amplified genes.


Author(s):  
Jeffrey T. Mcgrew ◽  
Cheryl L. Richards ◽  
Pauline Smidt ◽  
Bradley Dell ◽  
Virginia Price

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