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Provedor de dados:  Electron. J. Biotechnol.
País:  Chile
Título:  Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
Autores:  Fang,Qiangyi
Data:  2010-09-01
Ano:  2010
Palavras-chave:  Bioreactor
Human tissue kallikrein
Transient gene expression
Resumo:  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 µg/mL, a polyethylenimine concentration of 10.5 µg/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.
Tipo:  Journal article
Idioma:  Inglês
Editor:  Pontificia Universidad Católica de Valparaíso
Formato:  text/html
Fonte:  Electronic Journal of Biotechnology v.13 n.5 2010

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