Detailed understanding of enhanced specific productivity in Chinese hamster ovary cells at low culture temperature(CELL AND TISSUE ENGINEERING)
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概要
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The specific productivity of tumor necrosis factor receptor-immunoglobulin G1 Fc fusion (TNFR-Fc) (q_<TNFR-Fc>) in Chinese hamster ovary (CHO) cells at 30℃ was approximately 5-fold higher than that at 37℃. To investigate reasons for increased q_<TNFR-Fc> at low culture temperature, TNFR-Fc mRNA levels were determined by real-time PCR. It was found that like q_<TNFR-Fc>, the relative TNFR-Fc mRNA level was increased by lowering culture temperature, and more importantly, the kinetics of the increase in TNFR-Fc mRNA levels were in accordance with the changes in q_<TNFR-Fc>. The results demonstrated that the increased transcriptional level of TNFR-Fc was responsible for the increased q_<TNFR-Fc> at low culture temperature. Enhanced levels of mRNA could derive from increased gene copy number, improved mRNA stability, or enhanced transcriptional rate. There was not a big change of gene copy number by lowering culture temperature. The transcriptional rate of TNFR-Fc was slightly decreased at 30℃, compared to 37℃. However, mRNA stability of TNFR-Fc was significantly improved by lowering culture temperature. The half-life of TNFR-Fc mRNA was 5.55h at 30℃, whereas that was 3.69h at 37℃. Taken together, the reasons for the increased q_<TNFR-Fc> in CHO cells at low culture temperature were mainly the enhanced TNFR-Fc mRNA levels, which resulted from the improved mRNA stability, rather than the changes in the gene copy number or the transcriptional rate.
著者
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Ye Zhao-yang
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Kou Tian-Ci
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Fan Li
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Zhou Yan
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Liu Xu-Ping
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Zhao Liang
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Tan Wen-Song
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Tan Wen-song
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Liu Xu-ping
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Zhao Liang
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Zhou Yan
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Fan Li
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Kou Tian-ci
The State Key Laboratory Of Bioreactor Engineering East China University Of Science And Technology
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Koua Tian-Ci
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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Fana Li
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
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