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Nanoparticle-based capillary electroseparation of proteins in polymer capillaries under physiological conditions.

Nilsson, Christian LU ; Harwigsson, Ian; Becker, Kristian LU ; Kutter, Jörg P; Birnbaum, Staffan and Nilsson, Staffan LU (2010) In Electrophoresis 31(3). p.459-464
Abstract
Totally porous lipid-based liquid crystalline nanoparticles were used as pseudostationary phase for capillary electroseparation with LIF detection of proteins at physiological conditions using unmodified cyclic olefin copolymer capillaries (Topas, 6.7 cm effective length). In the absence of nanoparticles, i.e. in CE mode, the protein samples adsorbed completely to the capillary walls and could not be recovered. In contrast, nanoparticle-based capillary electroseparation resolved green fluorescent protein from several of its impurities within 1 min. Furthermore, a mixture of native green fluorescent protein and two of its single-amino-acid-substituted variants was separated within 2.5 min with efficiencies of 400 000 plates/m. The... (More)
Totally porous lipid-based liquid crystalline nanoparticles were used as pseudostationary phase for capillary electroseparation with LIF detection of proteins at physiological conditions using unmodified cyclic olefin copolymer capillaries (Topas, 6.7 cm effective length). In the absence of nanoparticles, i.e. in CE mode, the protein samples adsorbed completely to the capillary walls and could not be recovered. In contrast, nanoparticle-based capillary electroseparation resolved green fluorescent protein from several of its impurities within 1 min. Furthermore, a mixture of native green fluorescent protein and two of its single-amino-acid-substituted variants was separated within 2.5 min with efficiencies of 400 000 plates/m. The nanoparticles prevent adsorption by introducing a large interacting surface and by obstructing the attachment of the protein to the capillary wall. A one-step procedure based on self-assembly of lipids was used to prepare the nanoparticles, which benefit from their biocompatibility and suspension stability at high concentrations. An aqueous tricine buffer at pH 7.5 containing lipid-based nanoparticles (2% w/w) was used as electrolyte, enabling separation at protein friendly conditions. The developed capillary-based method facilitates future electrochromatography of proteins on polymer-based microchips under physiological conditions and enables the initial optimization of separation conditions in parallel to the chip development. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Electrophoresis
volume
31
issue
3
pages
459 - 464
publisher
John Wiley & Sons
external identifiers
  • wos:000274603200004
  • pmid:20119954
  • scopus:75749096659
ISSN
0173-0835
DOI
10.1002/elps.200900464
language
English
LU publication?
yes
id
225eb0f6-b736-4d7f-bc77-b7397b47478c (old id 1553117)
date added to LUP
2010-03-08 13:41:21
date last changed
2018-05-29 10:41:40
@article{225eb0f6-b736-4d7f-bc77-b7397b47478c,
  abstract     = {Totally porous lipid-based liquid crystalline nanoparticles were used as pseudostationary phase for capillary electroseparation with LIF detection of proteins at physiological conditions using unmodified cyclic olefin copolymer capillaries (Topas, 6.7 cm effective length). In the absence of nanoparticles, i.e. in CE mode, the protein samples adsorbed completely to the capillary walls and could not be recovered. In contrast, nanoparticle-based capillary electroseparation resolved green fluorescent protein from several of its impurities within 1 min. Furthermore, a mixture of native green fluorescent protein and two of its single-amino-acid-substituted variants was separated within 2.5 min with efficiencies of 400 000 plates/m. The nanoparticles prevent adsorption by introducing a large interacting surface and by obstructing the attachment of the protein to the capillary wall. A one-step procedure based on self-assembly of lipids was used to prepare the nanoparticles, which benefit from their biocompatibility and suspension stability at high concentrations. An aqueous tricine buffer at pH 7.5 containing lipid-based nanoparticles (2% w/w) was used as electrolyte, enabling separation at protein friendly conditions. The developed capillary-based method facilitates future electrochromatography of proteins on polymer-based microchips under physiological conditions and enables the initial optimization of separation conditions in parallel to the chip development.},
  author       = {Nilsson, Christian and Harwigsson, Ian and Becker, Kristian and Kutter, Jörg P and Birnbaum, Staffan and Nilsson, Staffan},
  issn         = {0173-0835},
  language     = {eng},
  number       = {3},
  pages        = {459--464},
  publisher    = {John Wiley & Sons},
  series       = {Electrophoresis},
  title        = {Nanoparticle-based capillary electroseparation of proteins in polymer capillaries under physiological conditions.},
  url          = {http://dx.doi.org/10.1002/elps.200900464},
  volume       = {31},
  year         = {2010},
}