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Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems.

Johansson, Hans-Olof LU ; Matos, Tiago LU ; Luz, Juliana S; Feitosa, Eloi; Oliveira, Carla C; Pessoa, Adalberto; Bülow, Leif LU and Tjerneld, Folke LU (2012) In Journal of Chromatography A 1233. p.30-35
Abstract
Phase diagrams of poly(ethylene glycol)/polyacrylate/Na(2)SO(4) systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coli can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coli homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more... (More)
Phase diagrams of poly(ethylene glycol)/polyacrylate/Na(2)SO(4) systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coli can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coli homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Na(2)SO(4)-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Chromatography A
volume
1233
pages
30 - 35
publisher
Elsevier
external identifiers
  • wos:000302663000005
  • pmid:22391492
  • scopus:84858622366
ISSN
0021-9673
DOI
10.1016/j.chroma.2012.02.028
language
English
LU publication?
yes
id
793e7e3d-a0e1-4d8a-9b34-32da3d3421c7 (old id 2432229)
date added to LUP
2012-04-12 17:17:40
date last changed
2017-03-05 03:50:38
@article{793e7e3d-a0e1-4d8a-9b34-32da3d3421c7,
  abstract     = {Phase diagrams of poly(ethylene glycol)/polyacrylate/Na(2)SO(4) systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coli can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coli homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Na(2)SO(4)-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH.},
  author       = {Johansson, Hans-Olof and Matos, Tiago and Luz, Juliana S and Feitosa, Eloi and Oliveira, Carla C and Pessoa, Adalberto and Bülow, Leif and Tjerneld, Folke},
  issn         = {0021-9673},
  language     = {eng},
  pages        = {30--35},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems.},
  url          = {http://dx.doi.org/10.1016/j.chroma.2012.02.028},
  volume       = {1233},
  year         = {2012},
}