title: Increasing the culture efficiency of hybridoma cells by the use of integrated metabolic control of glucose and glutamine at low levels.
abstract: The metabolism of HAb18 hybridoma cells was shifted to decrease metabolite accumulation and to improve culture efficiency by integrated metabolic control of glucose and glutamine at low levels. When glucose and glutamine levels were decreased to 0.5 and 0.3 mM respectively, lactate and ammonia production were reduced by 62.6 and 74% respectively, glucose-to-cell yield was increased from 0.23x10(9) to 0.66x10(9) cells.mmol-1, and glutamine-to-cell yield from 0.18x10(9) to 1.95x10(9) cells.mmol-1. Compared with high-level glucose and glutamine fed-batch cultures, low-level glucose and glutamine led to higher cell density (1.0x10(7) versus 0.3x10(7) cells.ml-1), longer culture span (14 as opposed to 8 days) and higher antibody yield (250 as against 150 mg.l-1). These results indicate that hybridoma culture efficiency would be increased by the integrated control of glucose and glutamine at 0.5 and 0.3 mM respectively. In contrast with previously reported glucose-and/or-glutamine-level-controlled fed-batch cultures, we demonstrated an efficient strategy of nutrient level selection and amino acid feeding. More importantly, our accurately and well-distributed Equable Feeding Control System opens a new avenue for reducing metabolites to low levels by controlling nutrients at low levels.
authors.affiliation: Cell Engineering Research Centre, State Key Laboratory of Cancer Biology, The Fourth Military Medical University, 17 Changle West Road, Xi'an 710032, People's Republic of China.
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Mi
authors.forename: Qiang
authors.initials: Q
authors.lastname: Feng
authors.forename: Rong
authors.initials: R
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Tang
authors.forename: Li
authors.initials: L
authors.lastname: Xie
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Yu
authors.forename: Zhinan
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000465:Algorithms; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D003198:Computer Simulation; D004734:Energy Metabolism / Q000502:physiology*; D005246:Feedback / Q000502:physiology; D005947:Glucose / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D006825:Hybridomas / Q000166:cytology* / Q000502:physiology*; D051379:Mice; D008954:Models, Biological
chemical_list: D005973:Glutamine; D005947:Glucose