Studies on optimal conditions of modified Immobilization carriers and additives for transgenic rice cell culture producing mGM-CSF
2010
Hochschulschrift
Zugriff:
98
Among the advantages offered by plants include the ease of scale-up with agriculture、reduced cost、the product safety as compared with microbial and mammalian production systems, and the capability of plants to correctly fold complex proteins. GM-CSF was expressed under the control of the sucrose starvation inducible rice αAmy3 promoter, and secretion into the culture medium was achieved by using the αAmy3 signal sequence.The transgenic rice cell, 1-10 (#14-4 for A1 intron) cell line with mGM-CSF production of 3.69 mg/L, was selected in this study. We generated mGM-CSF not only in suspension cultures,but also continued previous immobilization rice cell culture for deacylated loofa sponge.Finally,we evaluated the effects of additives on the production of recombinant target protein. In suspension cell culture, the best culture policy in 250 ml flask is high cellular density, N6 medium, auto induction, have the mGM-CSF production of 24.45 mg/L(day 22). In 2 L airlift culture, the highest production is 25.56 mg/L (day 25). In immobilization cell culture, we had the higher cell viability on both MS and N6 medium by elimination epidermis and deacylated loofa sponge in 1 L flask;and the highest production of 0.45 mg/L by elimination epidermis and deacylated of loofa sponge in N6 medium.At last, the mGM-CSF production of immobilization cell culture for elimination epidermis and deacylated loofa sponge in modified 2 L airlift is 1.53 mg/L (day 31). In order to stabilize the secreted mGM-CSF,two stabilizing polymers were tested (polyethylene glycol, and gelatin). Polyethylene glycol (PEG)was the most effective in stabilizing the secreted mGM-CSF. After the addition of 2% (w/v) PEG, the maximum mGM-CSF concentration reached 12.25 mg/L.In other way, the mGM-CSF production of add proline in N6 medium.After the addition of 4 flods proline(11.2 g-proline / L-N6),the maximum mGM-CSF concentration up to 36 mg/L.
Titel: |
Studies on optimal conditions of modified Immobilization carriers and additives for transgenic rice cell culture producing mGM-CSF
|
---|---|
Autor/in / Beteiligte Person: | Jyun Yu Liao ; 廖俊宇 |
Link: | |
Veröffentlichung: | 2010 |
Medientyp: | Hochschulschrift |
Sonstiges: |
|