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Plasmid adsorption to anion-exchange matrices: Comments on plasmid recovery

Tiainen, Peter LU ; Galaev, Igor LU and Larsson, Per-Olof LU (2007) In Biotechnology Journal 2(6). p.726-735
Abstract
A number of anion-exchange adsorbents were constructed, employing nonporous silica fibers, and examined with the aim of describing factors that influence desorption and recovery of plasmid DNA (pDNA). The fibers were provided with ligands via adsorption of the polymeric amines poly(ethyleneimine) or chitosan, or via graft-polymerization of primary, tertiary, or quaternary amine monomers to vinyl-silanized fibers. Several adsorbents showed an almost irreversible plasmid binding. It was suggested that important factors affecting the DNA releasing ability are (i) type of amine ligand used (primary amines bind plasmids the strongest), (ii) the structure of the nucleic acid (supercoiled pDNA may bind stronger than linear genomic DNA), (iii)... (More)
A number of anion-exchange adsorbents were constructed, employing nonporous silica fibers, and examined with the aim of describing factors that influence desorption and recovery of plasmid DNA (pDNA). The fibers were provided with ligands via adsorption of the polymeric amines poly(ethyleneimine) or chitosan, or via graft-polymerization of primary, tertiary, or quaternary amine monomers to vinyl-silanized fibers. Several adsorbents showed an almost irreversible plasmid binding. It was suggested that important factors affecting the DNA releasing ability are (i) type of amine ligand used (primary amines bind plasmids the strongest), (ii) the structure of the nucleic acid (supercoiled pDNA may bind stronger than linear genomic DNA), (iii) shift of ligand pKa (due to the proximity of highly charged pDNA), and (iv) the solid support itself (steric factors may lead to kinetically stable complexes). The last factor was derived from several comparisons between support-bound ligand and free soluble ligand. It was thus observed that polyelectrolyte complexes associated with a surface were much more difficult to dissociate than the equivalent soluble complexes. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Poly(ethyleneimine), Plasmid purification, Plasmid DNA, Anion-exchange chromatography, Recovery
in
Biotechnology Journal
volume
2
issue
6
pages
726 - 735
publisher
John Wiley & Sons
external identifiers
  • scopus:34447345938
ISSN
1860-6768
DOI
10.1002/biot.200700044
language
English
LU publication?
yes
id
02446ef9-046c-43a1-8c85-ec3a17a15ea8 (old id 792808)
date added to LUP
2008-01-08 17:26:19
date last changed
2017-01-01 05:00:15
@article{02446ef9-046c-43a1-8c85-ec3a17a15ea8,
  abstract     = {A number of anion-exchange adsorbents were constructed, employing nonporous silica fibers, and examined with the aim of describing factors that influence desorption and recovery of plasmid DNA (pDNA). The fibers were provided with ligands via adsorption of the polymeric amines poly(ethyleneimine) or chitosan, or via graft-polymerization of primary, tertiary, or quaternary amine monomers to vinyl-silanized fibers. Several adsorbents showed an almost irreversible plasmid binding. It was suggested that important factors affecting the DNA releasing ability are (i) type of amine ligand used (primary amines bind plasmids the strongest), (ii) the structure of the nucleic acid (supercoiled pDNA may bind stronger than linear genomic DNA), (iii) shift of ligand pKa (due to the proximity of highly charged pDNA), and (iv) the solid support itself (steric factors may lead to kinetically stable complexes). The last factor was derived from several comparisons between support-bound ligand and free soluble ligand. It was thus observed that polyelectrolyte complexes associated with a surface were much more difficult to dissociate than the equivalent soluble complexes.},
  author       = {Tiainen, Peter and Galaev, Igor and Larsson, Per-Olof},
  issn         = {1860-6768},
  keyword      = {Poly(ethyleneimine),Plasmid purification,Plasmid DNA,Anion-exchange chromatography,Recovery},
  language     = {eng},
  number       = {6},
  pages        = {726--735},
  publisher    = {John Wiley & Sons},
  series       = {Biotechnology Journal},
  title        = {Plasmid adsorption to anion-exchange matrices: Comments on plasmid recovery},
  url          = {http://dx.doi.org/10.1002/biot.200700044},
  volume       = {2},
  year         = {2007},
}