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Polyelectrolyte complexes as a tool for purification of plasmid DNA background and development

Wahlund, Per-Olof LU ; Gustavsson, Per-Erik LU ; Izumrudov, Vladimir LU ; Larsson, Per-Olof LU and Galaev, Igor LU (2004) In Journal of Chromatography. B 807(1). p.121-127
Abstract
The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Plasmid DNA was selectively precipitated from a clarified alkaline lysate using the polycation poly(N,N'-dimethyldiallylammonium) chloride which formed insoluble polyelectrolyte complex (PEC) with the plasmid DNA. Soluble PECs of DNA with polycations have earlier been used for cell transformation, but now the focus has been on insoluble PECs. Both DNA and RNA form stable PECs with synthetic polycations. However, it was possible to find a range of salt concentration where plasmid DNA was quantitatively precipitated whereas RNA remained in solution. The precipitated plasmid DNA was resolubilised... (More)
The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Plasmid DNA was selectively precipitated from a clarified alkaline lysate using the polycation poly(N,N'-dimethyldiallylammonium) chloride which formed insoluble polyelectrolyte complex (PEC) with the plasmid DNA. Soluble PECs of DNA with polycations have earlier been used for cell transformation, but now the focus has been on insoluble PECs. Both DNA and RNA form stable PECs with synthetic polycations. However, it was possible to find a range of salt concentration where plasmid DNA was quantitatively precipitated whereas RNA remained in solution. The precipitated plasmid DNA was resolubilised at high salt concentration and the polycation was removed by gel-filtration. (C) 2004 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Purification, Plasmid DNA, Polyelectrolyte complexes
in
Journal of Chromatography. B
volume
807
issue
1
pages
121 - 127
publisher
Elsevier
external identifiers
  • wos:000222047600018
  • pmid:15177169
  • scopus:2642571617
ISSN
1873-376X
DOI
10.1016/j.jchromb.2004.01.046
language
English
LU publication?
yes
id
5de49637-2174-4bd9-8909-e46de330a125 (old id 140658)
date added to LUP
2007-07-03 09:35:32
date last changed
2017-04-16 03:28:22
@article{5de49637-2174-4bd9-8909-e46de330a125,
  abstract     = {The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Plasmid DNA was selectively precipitated from a clarified alkaline lysate using the polycation poly(N,N'-dimethyldiallylammonium) chloride which formed insoluble polyelectrolyte complex (PEC) with the plasmid DNA. Soluble PECs of DNA with polycations have earlier been used for cell transformation, but now the focus has been on insoluble PECs. Both DNA and RNA form stable PECs with synthetic polycations. However, it was possible to find a range of salt concentration where plasmid DNA was quantitatively precipitated whereas RNA remained in solution. The precipitated plasmid DNA was resolubilised at high salt concentration and the polycation was removed by gel-filtration. (C) 2004 Elsevier B.V. All rights reserved.},
  author       = {Wahlund, Per-Olof and Gustavsson, Per-Erik and Izumrudov, Vladimir and Larsson, Per-Olof and Galaev, Igor},
  issn         = {1873-376X},
  keyword      = {Purification,Plasmid DNA,Polyelectrolyte complexes},
  language     = {eng},
  number       = {1},
  pages        = {121--127},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography. B},
  title        = {Polyelectrolyte complexes as a tool for purification of plasmid DNA background and development},
  url          = {http://dx.doi.org/10.1016/j.jchromb.2004.01.046},
  volume       = {807},
  year         = {2004},
}