Overexpression of GADD34 enhances production of recombinant human antithrombin III in Chinese hamster ovary cells.
To improve the production of recombinant human antithrombin III (AT-III) in Chinese hamster ovary (CHO) cells, the gene encoding growth arrest and DNA damage inducible protein 34 (GADD34), which is a transcription factor involved in the unfolded protein response (UPR), was cloned from CHO-K1 cells. Overexpression of GADD34 significantly enhanced the production of recombinant AT-III in CHO 13D-35D cells. The specific rate of AT-III production in the GADD34-overexpressing CHO 13D-35D cells reached approximately 28 pg/cell/d. After 144 h of incubation, the AT-III concentration in the culture supernatant was approximately 40% higher than that observed in the case of the parental CHO 13D-35D cells. The mRNA expression, specific activity, and fucosylation of AT-III were not affected by GADD34 overexpression. Overexpression of GADD34 is a promising method of improving the production of secreted protein pharmaceuticals in CHO cells.
Omasa
Takeshi
T
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan. omasa@bio.eng.osaka-u.ac.jp
Takami
Takashi
T
Ohya
Tomoshi
T
Kiyama
Eriko
E
Hayashi
Tetsuji
T
Nishii
Hiroko
H
Miki
Hideo
H
Kobayashi
Kaoru
K
Honda
Kohsuke
K
Ohtake
Hisao
H
D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics*; D000990:Antithrombin III / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D016466:CHO Cells; D018797:Cell Cycle Proteins / Q000235:genetics*; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D006801:Humans; D054645:Protein Phosphatase 1; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D000943:Antigens, Differentiation; D018797:Cell Cycle Proteins; D017931:DNA Primers; D011994:Recombinant Proteins; D000990:Antithrombin III; C087519:PPP1R15A protein, human; D054645:Protein Phosphatase 1