GROWTH-CHARACTERISTICS IN FED-BATCH CULTURE OF HYBRIDOMA CELLS WITH CONTROL OF GLUCOSE AND GLUTAMINE CONCENTRATIONS
GROWTH-CHARACTERISTICS IN FED-BATCH CULTURE OF HYBRIDOMA CELLS WITH CONTROL OF GLUCOSE AND GLUTAMINE CONCENTRATIONS
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DOI：
10.1002/bit.260440114
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发表时间：
1994-06-05
影响因子：
3.8
通讯作者：
KOBAYASHI, T
中科院分区：
文献类型：
--
作者：
KUROKAWA, H;PARK, YS;KOBAYASHI, T
开发了一种利用高效液相色谱在线测定发酵液中葡萄糖、谷氨酰胺和乳酸的系统。利用该系统，采用自适应控制算法，将葡萄糖和谷氨酰胺的浓度分别控制在0.2~2.0和0.1~0.6g/L范围内。当葡萄糖浓度控制在0.2g/L时，胞内乳酸脱氢酶活性下降了一半，乳酸浓度下降了三分之一，丝氨酸和甘氨酸的吸收速率大约是控制在1.0g/L时的两倍，而当葡萄糖浓度较低时，氨的产生增加了。在杂交瘤细胞培养中，为了减少氨和乳酸等抑制代谢产物的产生，提高抗体产生率，葡萄糖和谷氨酰胺的浓度分别控制在0.2g/L和0.1g/100g/g。在这些低浓度的葡萄糖和谷氨酰胺的作用下，细胞浓度(4.1x10(6)个/毫升)和抗体产生(172 mg/L)都比控制较高的葡萄糖水平时增加了大约两倍。根据这些结果，同时控制葡萄糖和谷氨酰胺的浓度对于杂交瘤细胞培养产生抗体是有用的。(C)1994年John Wiley父子公司。
An online system using HPLC was developed for the measurement of glucose, glutamine, and lactate in a culture broth. Using the system, the glucose and glutamine concentrations were controlled simultaneously by an adaptive-control algorithm within the ranges of 0.2 to 2.0 and 0.1 to 0.6 g/L, respectively. When the glucose concentration was controlled at the low level of 0.2 g/L, the intracellular lactate dehydrogenase activity decreased by one-half and the lactate concentration by one-third, whereas the uptake rates of serine and glycine were about twice as high, compared with the amounts when the glucose concentration was control led at 1.0 g/L. On the other hand, ammonia production increased when the glucose concentration was kept low. To reduce the production of inhibitory metabolites such as ammonia and lactate and improve the antibody production rate in a hybridoma cell culture, the concentrations of glucose and glutamine were controlled at 0.2 and 0.1 g/L, respectively. With these low concentrations of glucose and glutamine, the cell concentration (4.1 x 10(6) cells/mL) and antibody production (172 mg/L) both increased about twofold compared with the amounts when the glucose was controlled at higher levels. From these results, simultaneous control of the glucose and glutamine concentrations was shown to be useful in the production of antibody by hybridoma cell cultivation. (C) 1994 John Wiley and Sons, Inc.