Optimization of polyethylenimine-mediated transient transfection using response surface methodology design

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 a...

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Autores principales:	Fang,Qiangyi, Shen,Bingqian
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	bioreactor human tissue kallikrein transient gene expression
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500010
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spelling	oai:scielo:S0717-345820100005000102011-05-24Optimization of polyethylenimine-mediated transient transfection using response surface methodology designFang,QiangyiShen,Bingqian bioreactor human tissue kallikrein transient gene expression 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.5 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500010en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	bioreactor human tissue kallikrein transient gene expression
spellingShingle	bioreactor human tissue kallikrein transient gene expression Fang,Qiangyi Shen,Bingqian Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
description	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.
author	Fang,Qiangyi Shen,Bingqian
author_facet	Fang,Qiangyi Shen,Bingqian
author_sort	Fang,Qiangyi
title	Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
title_short	Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
title_full	Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
title_fullStr	Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
title_full_unstemmed	Optimization of polyethylenimine-mediated transient transfection using response surface methodology design
title_sort	optimization of polyethylenimine-mediated transient transfection using response surface methodology design
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500010
work_keys_str_mv	AT fangqiangyi optimizationofpolyethyleniminemediatedtransienttransfectionusingresponsesurfacemethodologydesign AT shenbingqian optimizationofpolyethyleniminemediatedtransienttransfectionusingresponsesurfacemethodologydesign
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Optimization of polyethylenimine-mediated transient transfection using response surface methodology design

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