Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
Electron. j. biotechnol
;
13(5): 10-11, Sept. 2010. ilus, tab
Article
in English
| LILACS
| ID: lil-591892
ABSTRACT
Response surface methodology was undertaken to optimize the polyethylenimine-mediated transient transfection of suspension cultured HEK 293-F cells. A total of 15 combinations were designed according to Box-Behnken design to identify the effects of DNA concentration, polyethylenimine concentration and incubation time on transient transfection efficiency. The highest integral optic density of green fluorescent protein presenting r-protein yield was accessed using a DNA concentration of 1.75 ug/mL, a polyethylenimine concentration of 10.5 ug/mL, and an incubation time of 11.8 min. Analysis of variance demonstrated that the experimental values fit well with a quadratic model. The RSM-optimized transient transfection resulted in greater production of human tissue prokallikrein (TproK) than non-RSM-optimized conditions protein yield was 32.0 mg/L and the maximum viable cell density reached 3.57 x 10(6) cells/mL in a 5 L stirred-tank bioreactor culture.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Transfection
/
Gene Expression
/
Tissue Kallikreins
Limits:
Humans
Language:
English
Journal:
Electron. j. biotechnol
Journal subject:
Biotechnology
Year:
2010
Type:
Article
Affiliation country:
China
Institution/Affiliation country:
Chinese Academy of Sciences/CN
/
Nankai University/CN
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