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Plasmid purification using non-porous anion-exchange silica fibres

Tiainen, Peter LU ; Gustavsson, Per-Erik LU ; Månsson, Mats-Olle LU and Larsson, Per-Olof LU (2007) In Journal of Chromatography A 1149(2). p.158-168
Abstract
A new type of fibre-based anion-exchange material for plasmid purification was developed. The basic material consisted of non-porous silica fibres with a mean diameter of 1.5 mu m and a surface area of 2.4 m(2) g(-1). The fibre surface was provided with several types of ligands, either by adsorption of polymers (chitosan or poly(ethyleneinune)) or by polymerization of amine-containing acrylic monomers onto a propyl methacrylatesilanized surface. The resulting polymer layers contained primary, tertiary or quaternary amines as ion-exchange groups. The packing density could be varied considerably, 9-34% (v/v). The loose packing structure provided excellent flow properties suitable for high-speed operations. The best overall performance was... (More)
A new type of fibre-based anion-exchange material for plasmid purification was developed. The basic material consisted of non-porous silica fibres with a mean diameter of 1.5 mu m and a surface area of 2.4 m(2) g(-1). The fibre surface was provided with several types of ligands, either by adsorption of polymers (chitosan or poly(ethyleneinune)) or by polymerization of amine-containing acrylic monomers onto a propyl methacrylatesilanized surface. The resulting polymer layers contained primary, tertiary or quaternary amines as ion-exchange groups. The packing density could be varied considerably, 9-34% (v/v). The loose packing structure provided excellent flow properties suitable for high-speed operations. The best overall performance was shown by silica fibres provided with tertiary amine polymers, having a plasmid-binding capacity of 0.9 mg ml(-1) (pre-purified plasmid) and a plasmid recovery of 62% (performance data remained stable though several adsorption cycles). The high flow rates possible with the fibre material made it especially useful when large volumes of cleared lysate were processed. The columns could be operated with retention of their adsorption properties at speeds of up to 1800 cm h(-1) equivalent to 0.5 column volumes per minute. The binding capacity was found to be lower than anticipated from the design of the fibres. Fluorescence imaging showing individual plasmid molecules indicated the fibre population to be heterogeneous with respect to plasmid adsorption, some fibres displaying poor binding properties. Possible reasons for this heterogeneity are discussed. (c) 2007 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
plasmid, silica fibres, anion-exchange chromatography, plasmid DNA, plasmid purification, imaging
in
Journal of Chromatography A
volume
1149
issue
2
pages
158 - 168
publisher
Elsevier
external identifiers
  • wos:000246673000005
  • scopus:34247190615
ISSN
0021-9673
DOI
10.1016/j.chroma.2007.02.114
language
English
LU publication?
yes
id
c42f0ac1-486f-4e04-8592-8f2c53552d73 (old id 659452)
date added to LUP
2007-12-19 09:30:51
date last changed
2017-01-01 07:15:44
@article{c42f0ac1-486f-4e04-8592-8f2c53552d73,
  abstract     = {A new type of fibre-based anion-exchange material for plasmid purification was developed. The basic material consisted of non-porous silica fibres with a mean diameter of 1.5 mu m and a surface area of 2.4 m(2) g(-1). The fibre surface was provided with several types of ligands, either by adsorption of polymers (chitosan or poly(ethyleneinune)) or by polymerization of amine-containing acrylic monomers onto a propyl methacrylatesilanized surface. The resulting polymer layers contained primary, tertiary or quaternary amines as ion-exchange groups. The packing density could be varied considerably, 9-34% (v/v). The loose packing structure provided excellent flow properties suitable for high-speed operations. The best overall performance was shown by silica fibres provided with tertiary amine polymers, having a plasmid-binding capacity of 0.9 mg ml(-1) (pre-purified plasmid) and a plasmid recovery of 62% (performance data remained stable though several adsorption cycles). The high flow rates possible with the fibre material made it especially useful when large volumes of cleared lysate were processed. The columns could be operated with retention of their adsorption properties at speeds of up to 1800 cm h(-1) equivalent to 0.5 column volumes per minute. The binding capacity was found to be lower than anticipated from the design of the fibres. Fluorescence imaging showing individual plasmid molecules indicated the fibre population to be heterogeneous with respect to plasmid adsorption, some fibres displaying poor binding properties. Possible reasons for this heterogeneity are discussed. (c) 2007 Elsevier B.V. All rights reserved.},
  author       = {Tiainen, Peter and Gustavsson, Per-Erik and Månsson, Mats-Olle and Larsson, Per-Olof},
  issn         = {0021-9673},
  keyword      = {plasmid,silica fibres,anion-exchange chromatography,plasmid DNA,plasmid purification,imaging},
  language     = {eng},
  number       = {2},
  pages        = {158--168},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Plasmid purification using non-porous anion-exchange silica fibres},
  url          = {http://dx.doi.org/10.1016/j.chroma.2007.02.114},
  volume       = {1149},
  year         = {2007},
}