| RRC ID |
43202
|
| 著者 |
Ohya T, Hayashi T, Kiyama E, Nishii H, Miki H, Kobayashi K, Honda K, Omasa T, Ohtake H.
|
| タイトル |
Improved production of recombinant human antithrombin III in Chinese hamster ovary cells by ATF4 overexpression.
|
| ジャーナル |
Biotechnol Bioeng
|
| Abstract |
To improve the production of recombinant human antithrombin III (AT-III) in Chinese hamster ovary (CHO) cells, the genes encoding transcription factors, ATF4 (activating transcription factor 4) and XBP-1s (the spliced form of X-box binding protein 1), which were involved in the mammalian unfolded protein response (UPR), were cloned from CHO-K1 cells. Overexpression of ATF4 significantly enhanced the production of recombinant AT-III in CHO 13D-35D cells. The specific rate of AT-III production in the ATF4-overexpressed CHO 13D-35D cells reached approximately 23 pg/cell/day. After 144 h of incubation, the AT-III concentration in the culture supernatant was twofold greater compared to that observed with parental CHO 13D-35D cells. In contrast, ectopic expression of XBP-1s failed to enhance the production of recombinant AT-III in CHO 13D-35D cells. RT-PCR analysis revealed that high levels of XBP-1s mRNA were present in the CHO cells, regardless of ectopic expression of XBP-1s. Our results indicate that overexpression of the UPR transcription factor ATF4 is a promising means for improving the production of secreted protein pharmaceuticals in CHO cells.
|
| 巻・号 |
100(2)
|
| ページ |
317-24
|
| 公開日 |
2008-6-1
|
| DOI |
10.1002/bit.21758
|
| PMID |
18078289
|
| MeSH |
Activating Transcription Factor 4 / genetics
Activating Transcription Factor 4 / metabolism*
Animals
Antithrombin III / biosynthesis*
Antithrombin III / genetics
CHO Cells / metabolism*
Cricetinae
Cricetulus
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Genetic Enhancement / methods*
Humans
Nuclear Proteins / genetics
Nuclear Proteins / metabolism*
Protein Engineering / methods*
Recombinant Proteins / biosynthesis
Regulatory Factor X Transcription Factors
Transcription Factors
X-Box Binding Protein 1
|
| IF |
4.002
|
| 引用数 |
70
|
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
|
