Hydrophobic Interaction Capillary Electrochromatography of Protein Mutants. Use of Lipid-Based Liquid Crystalline Nanoparticles as Pseudostationary Phase.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1276573
- author
- Nilsson, Christian LU ; Becker, Kristian LU ; Harwigsson, Ian ; Bülow, Leif LU ; Birnbaum, Staffan and Nilsson, Staffan LU
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Chemistry
- volume
- 81
- issue
- 1
- pages
- 315 - 321
- publisher
- The American Chemical Society (ACS)
- 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
- 2016-04-01 11:32:53
- date last changed
- 2022-01-26 06:57:59
@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 (ACS)}},
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}},
doi = {{10.1021/ac8020533}},
volume = {{81}},
year = {{2009}},
}