Li Ling, Mi Li, Feng Qiang, Liu Rong, Tang Hao, Xie Li, Yu Xiaoling, Chen Zhinan
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.
Biotechnol Appl Biochem. 2005 Aug;42(Pt 1):73-80. doi: 10.1042/BA20040203.
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.
通过在低水平对葡萄糖和谷氨酰胺进行综合代谢控制，HAb18杂交瘤细胞的代谢发生转变，以减少代谢产物积累并提高培养效率。当葡萄糖和谷氨酰胺水平分别降至0.5和0.3 mM时，乳酸和氨的产量分别降低了62.6%和74%，葡萄糖生成细胞的产量从0.23×10⁹增加到0.66×10⁹个细胞·mmol⁻¹，谷氨酰胺生成细胞的产量从0.18×10⁹增加到1.95×10⁹个细胞·mmol⁻¹。与高糖高谷氨酰胺补料分批培养相比，低糖低谷氨酰胺培养导致更高的细胞密度（1.0×10⁷对0.3×10⁷个细胞·ml⁻¹）、更长的培养时间（14天而非8天）和更高的抗体产量（250对150 mg·l⁻¹）。这些结果表明，分别将葡萄糖和谷氨酰胺控制在0.5和0.3 mM进行综合控制可提高杂交瘤培养效率。与先前报道的葡萄糖和/或谷氨酰胺水平控制的补料分批培养不同，我们展示了一种有效的营养水平选择和氨基酸补料策略。更重要的是，我们精确且分布良好的均衡补料控制系统通过在低水平控制营养物质为将代谢产物降至低水平开辟了一条新途径。
