title: Effect of low culture temperature on specific productivity and transcription level of anti-4-1BB antibody in recombinant Chinese hamster ovary cells.
abstract: Lowering the culture temperature has been suggested as a useful tool for improving the production of recombinant proteins in Chinese hamster ovary (CHO) cells. In an effort to improve anti-4-1BB antibody production in recombinant CHO (rCHO) cells, rCHO cells producing anti-4-1BB antibody (LGA31-56) were cultivated at three different temperatures, 30, 33, and 37 degrees C. Lowering the culture temperature led to suppressed cell growth, cell cycle arrest in G(0)/G(1) phase, and improved cell viability for a longer period. However, antibody production and q(Ab) were not increased at low culture temperature. The maximum antibody concentration and q(Ab) at 37 degrees C were 110.6 +/- 2.6 microg mL(-)(1) and 0.43 +/- 0.03 microg (10(6) cells h)(-)(1), respectively, whereas those at 30 degrees C were 28.3 +/- 3.8 microg mL(-)(1) and 0.44 +/- 0.07 (10(6) cells h)(-)(1), respectively. Northern blot analysis revealed that lowering the culture temperature did not increase the transcription level of heavy and light chains. These results were quite in contrast with the improved production of erythropoietin, which is expressed in the same CHO host and driven by the same CMV promoters, by lowering the temperature. Taken together, the results obtained imply that the beneficial effect of low culture temperature on recombinant protein production in rCHO cells is cell-line-specific.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 371-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea.
authors.forename: Sung Kwan
authors.initials: SK
authors.lastname: Yoon
authors.forename: Sung Hyun
authors.initials: SH
authors.lastname: Kim
authors.forename: Gyun Min
authors.initials: GM
authors.lastname: Lee
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism* / Q000528:radiation effects*; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000528:radiation effects; D002470:Cell Survival / Q000528:radiation effects; D003080:Cold Temperature; D006224:Cricetinae; D003412:Cricetulus; D004307:Dose-Response Relationship, Radiation; D005786:Gene Expression Regulation / Q000528:radiation effects; D006801:Humans; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013696:Temperature; D014158:Transcription, Genetic / Q000502:physiology* / Q000528:radiation effects*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins