Data/hora: 12/06/2024 10:09:01

Provedor de dados: | 69
País: | Chile
Título: | Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
Autores: | Fang,Qiangyi; Shen,Bingqian.
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
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500010
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.13 n.5 2010