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Purification of plasmid DNA with polymer-salt aqueous two-phase system: Optimization using response surface methodology

Rahimpour, Farshad ; Feyzi, Farzaneh ; Maghsoudi, Saeid and Hatti-Kaul, Rajni LU (2006) In Biotechnology and Bioengineering 95(4). p.627-637
Abstract
An experimental design was used to optimize plasmid purification from an alkaline lysate of Escherichia coli cells using PEG-sodium citrate aqueous two-phase systems (ATIPS), and to evaluate the influence of pH, PEG molecular weight, tie line length, phase volume ratio, and lysate load. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology (RMS) with an orthogonal rotatable central composite design was defined based on the conditions found for the highest purification by preliminary tests. The adequacy of the calculated models for the plasmid recovery and remaining RNA were confirmed by means of variance analysis and additional experiments. Analysis of contours of... (More)
An experimental design was used to optimize plasmid purification from an alkaline lysate of Escherichia coli cells using PEG-sodium citrate aqueous two-phase systems (ATIPS), and to evaluate the influence of pH, PEG molecular weight, tie line length, phase volume ratio, and lysate load. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology (RMS) with an orthogonal rotatable central composite design was defined based on the conditions found for the highest purification by preliminary tests. The adequacy of the calculated models for the plasmid recovery and remaining RNA were confirmed by means of variance analysis and additional experiments. Analysis of contours of constant response as a function of pH, PEG molecular weight, tie line length, and cell lysate load for three different phase volume ratios revealed different effects of these five factors on the studied parameters. Plasmid recovery of 99% was predicted for a system with PEG 400, pH 6.9, tie line length of 38.7%, phase volume ratio of 1.5, and lysate load of 10% MO. Under these conditions the predicted RNA removal was 68%. (c) 2006 Wiley Periodicals, Inc. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
central composition design, plasmid DNA purification, PEG-citrate aqueous two-phase systems, RNA removal
in
Biotechnology and Bioengineering
volume
95
issue
4
pages
627 - 637
publisher
John Wiley and Sons
external identifiers
  • wos:000241264300007
  • scopus:33750590040
ISSN
1097-0290
DOI
10.1002/bit.20920
language
English
LU publication?
yes
id
4f5a450c-bee5-4c33-ab81-554f673063f5 (old id 388210)
date added to LUP
2016-04-01 12:10:34
date last changed
2021-05-26 02:29:27
@article{4f5a450c-bee5-4c33-ab81-554f673063f5,
  abstract     = {An experimental design was used to optimize plasmid purification from an alkaline lysate of Escherichia coli cells using PEG-sodium citrate aqueous two-phase systems (ATIPS), and to evaluate the influence of pH, PEG molecular weight, tie line length, phase volume ratio, and lysate load. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology (RMS) with an orthogonal rotatable central composite design was defined based on the conditions found for the highest purification by preliminary tests. The adequacy of the calculated models for the plasmid recovery and remaining RNA were confirmed by means of variance analysis and additional experiments. Analysis of contours of constant response as a function of pH, PEG molecular weight, tie line length, and cell lysate load for three different phase volume ratios revealed different effects of these five factors on the studied parameters. Plasmid recovery of 99% was predicted for a system with PEG 400, pH 6.9, tie line length of 38.7%, phase volume ratio of 1.5, and lysate load of 10% MO. Under these conditions the predicted RNA removal was 68%. (c) 2006 Wiley Periodicals, Inc.},
  author       = {Rahimpour, Farshad and Feyzi, Farzaneh and Maghsoudi, Saeid and Hatti-Kaul, Rajni},
  issn         = {1097-0290},
  language     = {eng},
  number       = {4},
  pages        = {627--637},
  publisher    = {John Wiley and Sons},
  series       = {Biotechnology and Bioengineering},
  title        = {Purification of plasmid DNA with polymer-salt aqueous two-phase system: Optimization using response surface methodology},
  url          = {http://dx.doi.org/10.1002/bit.20920},
  doi          = {10.1002/bit.20920},
  volume       = {95},
  year         = {2006},
}