Development of apoptosis-resistant dihydrofolate reductase-deficient Chinese hamster ovary cell line.
Apoptosis-resistant dihydrofolate reductase-deficient CHO cell line (dhfr(-) CHO-bcl2) was developed by introduction of the bcl-2 gene into the dhfr(-) CHO cell line (DUKX-B11, ATCC CRL-9096) and subsequent selection of clones stably overexpressing Bcl-2 in the absence of selection pressure. When the dhfr(-) CHO-bcl2 cell line was used as a host cell line for development of a recombinant CHO (rCHO) cell line expressing a humanized antibody, it displayed stable expression of the bcl-2 gene during rCHO cell line development and no detrimental effect of Bcl-2 overexpression on specific antibody productivity. Taken together, the results obtained demonstrate that the use of an apoptosis-resistant dhfr(-) CHO cell line as the host cell line saves the effort of establishing an apoptosis-resistant rCHO cell line and expedites the development process of apoptosis-resistant rCHO cells producing therapeutic proteins.
Lee
Suk Kyoo
SK
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea.
Lee
Gyun Min
GM
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002074:Butanones / Q000494:pharmacology; D016466:CHO Cells / Q000145:classification* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005240:Feasibility Studies; D005784:Gene Amplification; D019254:Genes, bcl-2 / Q000235:genetics; D006801:Humans; D008727:Methotrexate / Q000494:pharmacology; D000077430:Nabumetone; D015202:Protein Engineering / Q000379:methods; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013045:Species Specificity; D013762:Tetrahydrofolate Dehydrogenase / Q000172:deficiency*
D000906:Antibodies; D002074:Butanones; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D013762:Tetrahydrofolate Dehydrogenase; D000077430:Nabumetone; D008727:Methotrexate