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首页> 外文期刊>Biotechnology Progress >Effect of Low Culture Temperature on Specific Productivity and Transcription Level of Anti-4-1BB Antibody in Recombinant Chinese Hamster Ovary Cells
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Effect of Low Culture Temperature on Specific Productivity and Transcription Level of Anti-4-1BB Antibody in Recombinant Chinese Hamster Ovary Cells

机译：低温培养对重组中国仓鼠卵巢细胞抗4-1BB抗体特异性生产力和转录水平的影响

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摘要

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 prductio in recombinant CHO (rCHO) cells, rCHO cells prdoucing anti-3-1BB antibody (LGA31-56) were cultivated at three different temperatures, 30, 33, and 37 deg 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 qAb were not increased at low culture temperature. The maximum antibody concentration and qAb at 37 deg C were 110.6 (+-) 2.6 mug mL~(-1) and 0.43 (+-) 0.03 mug (10~6 cells h)~(-1), respectively, whereas those at 30 deg C were 28.3 (+-) 3.8 mug 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 i nthe 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.
机译：已经提出降低培养温度是改善中国仓鼠卵巢（CHO）细胞中重组蛋白产生的有用工具。为了提高重组CHO（rCHO）细胞中抗4-1BB抗体的水平，在30、33和37℃的三个不同温度下培养了抗3-1BB抗体（LGA31-56）的rCHO细胞。降低培养温度可导致细胞生长受到抑制，细胞周期停滞在G_0 / G_1期，并在更长的时间内提高了细胞活力。但是，在低培养温度下抗体产量和qAb并未增加。在37摄氏度时的最大抗体浓度和qAb分别为110.6（+-）2.6杯mL〜（-1）和0.43（+-）0.03杯（10〜6个细胞h）〜（-1），而在30℃分别为28.3（±）3.8毫升mL〜（-1）和0.44（±）0.07（10〜6电池小时）〜（-1）。 Northern印迹分析表明，降低培养温度不会增加重链和轻链的转录水平。这些结果与促红细胞生成素产生的改善形成对照，促红细胞生成素通过降低温度在相同的CHO宿主中表达，并由相同的CMV启动子驱动。两者合计，获得的结果暗示低培养温度对rCHO细胞中重组蛋白产生的有益作用是细胞系特异性的。

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