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Hydrophobic Interaction Capillary Electrochromatography of Protein Mutants. Use of Lipid-Based Liquid Crystalline Nanoparticles as Pseudostationary Phase.

Nilsson, Christian LU ; Becker, Kristian LU ; Harwigsson, Ian; Bülow, Leif LU ; Birnbaum, Staffan and Nilsson, Staffan LU (2009) In Analytical Chemistry 81(1). p.315-321
Abstract
Nanoparticle-based hydrophobic interaction-capillary electrochromatography was utilized for separation of proteins with similar mass-to-charge ratio at neutral pH without organic modifier. Lipid-based liquid crystalline nanoparticles were prepared and used as pseudostationary phase, benefiting from their high biocompatibility, ease of preparation, and suspension stability at high concentrations. Use of laser-induced fluorescence enabled detection at high nanoparticle concentrations. Green fluorescent protein (GFP) and mutants of GFP harboring single or double amino acid substitutions with the same charge were separated in the described system but not in conventional capillary electrophoresis. Separation was achieved by increasing the salt... (More)
Nanoparticle-based hydrophobic interaction-capillary electrochromatography was utilized for separation of proteins with similar mass-to-charge ratio at neutral pH without organic modifier. Lipid-based liquid crystalline nanoparticles were prepared and used as pseudostationary phase, benefiting from their high biocompatibility, ease of preparation, and suspension stability at high concentrations. Use of laser-induced fluorescence enabled detection at high nanoparticle concentrations. Green fluorescent protein (GFP) and mutants of GFP harboring single or double amino acid substitutions with the same charge were separated in the described system but not in conventional capillary electrophoresis. Separation was achieved by increasing the salt concentration to promote hydrophobic interactions by shielding of the repulsive electrostatic interactions. In addition, the method was adapted to a capillary with an effective length of 6.7 cm, enabling fast separations and future applications on chip. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
81
issue
1
pages
315 - 321
publisher
The American Chemical Society
external identifiers
  • wos:000262113400045
  • pmid:19049363
  • scopus:58149468835
ISSN
1520-6882
DOI
10.1021/ac8020533
language
English
LU publication?
yes
id
71695200-de56-4b35-a504-20e05168cfe9 (old id 1276573)
date added to LUP
2009-01-27 16:53:00
date last changed
2017-10-22 03:32:45
@article{71695200-de56-4b35-a504-20e05168cfe9,
  abstract     = {Nanoparticle-based hydrophobic interaction-capillary electrochromatography was utilized for separation of proteins with similar mass-to-charge ratio at neutral pH without organic modifier. Lipid-based liquid crystalline nanoparticles were prepared and used as pseudostationary phase, benefiting from their high biocompatibility, ease of preparation, and suspension stability at high concentrations. Use of laser-induced fluorescence enabled detection at high nanoparticle concentrations. Green fluorescent protein (GFP) and mutants of GFP harboring single or double amino acid substitutions with the same charge were separated in the described system but not in conventional capillary electrophoresis. Separation was achieved by increasing the salt concentration to promote hydrophobic interactions by shielding of the repulsive electrostatic interactions. In addition, the method was adapted to a capillary with an effective length of 6.7 cm, enabling fast separations and future applications on chip.},
  author       = {Nilsson, Christian and Becker, Kristian and Harwigsson, Ian and Bülow, Leif and Birnbaum, Staffan and Nilsson, Staffan},
  issn         = {1520-6882},
  language     = {eng},
  number       = {1},
  pages        = {315--321},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Hydrophobic Interaction Capillary Electrochromatography of Protein Mutants. Use of Lipid-Based Liquid Crystalline Nanoparticles as Pseudostationary Phase.},
  url          = {http://dx.doi.org/10.1021/ac8020533},
  volume       = {81},
  year         = {2009},
}