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Nanoparticle-based capillary electrochromatography

Mannaa, Atef LU (2013) KEMP36 20131
Department of Chemistry
Abstract
In this work, lipid nanoparticles were used as pseudostationary phase, PSP, for protein separation in capillary electrochromatography, CEC. The aim of the work was to use a new analytical separation technique for future analysis of protein drugs by using lipid nanoparticles as a pseudo-stationary phase with conventional UV-detection.
PSP-CEC was combined with UV-detection to study proteins elution. The tested proteins BSA, cytochrome c and insulin have been selected according to different hydrophobicities, sizes and isoelectric points (pI). A zwitter ionic buffer, MOPS, was used as an electrolyte at pH 7.4. The nanoparticles were tested for their compatibility with MALDI-TOF/MS (Matrix assisted laser desorption/ionization-time of flight).... (More)
In this work, lipid nanoparticles were used as pseudostationary phase, PSP, for protein separation in capillary electrochromatography, CEC. The aim of the work was to use a new analytical separation technique for future analysis of protein drugs by using lipid nanoparticles as a pseudo-stationary phase with conventional UV-detection.
PSP-CEC was combined with UV-detection to study proteins elution. The tested proteins BSA, cytochrome c and insulin have been selected according to different hydrophobicities, sizes and isoelectric points (pI). A zwitter ionic buffer, MOPS, was used as an electrolyte at pH 7.4. The nanoparticles were tested for their compatibility with MALDI-TOF/MS (Matrix assisted laser desorption/ionization-time of flight). MADI-TOF/MS works with a suspension of lipid nanoparticles in MOPS that contains insulin and it can be a future possible detection method for PSP-CEC. All three proteins were successfully eluted with PSP-CEC combined with UV-detection. Nanoparticles coated with silver might be studied with “surface enhanced Raman spectroscopy”-detection. (Less)
Popular Abstract
Popular Science Summery
Capillary electrophoresis, CE is a separation technique in which analytes are separated according to their size to charge ratio in the interior of the capillary.
Capillary electrochromatography, CEC is a combination of CE and HPLC. The fundamental difference between CE and CEC is that the separation is not based on the electrophoretic mobility difference only but on the interaction with the stationary phase as well. Lipid nanoparticles were used as pseudostationary phase (moving phase). The aim of current work is to separate different proteins, BSA, cytochrome c and insulin have been separated according to their charge and size. They have been successfully separated. It was found that PSP-CEC in combination with... (More)
Popular Science Summery
Capillary electrophoresis, CE is a separation technique in which analytes are separated according to their size to charge ratio in the interior of the capillary.
Capillary electrochromatography, CEC is a combination of CE and HPLC. The fundamental difference between CE and CEC is that the separation is not based on the electrophoretic mobility difference only but on the interaction with the stationary phase as well. Lipid nanoparticles were used as pseudostationary phase (moving phase). The aim of current work is to separate different proteins, BSA, cytochrome c and insulin have been separated according to their charge and size. They have been successfully separated. It was found that PSP-CEC in combination with UV-detection is a good methodology to elute Proteins. An identification technique named MALDI-TOF/MS was used to identify the insulin that has been dissolved in nanoparticles. Insulin has been successfully identified. MADI-TOF/MS works well with lipid nanoparticles and it can be used as a future possible detection of PSP-CEC. (Less)
Please use this url to cite or link to this publication:
author
Mannaa, Atef LU
supervisor
organization
course
KEMP36 20131
year
type
L3 - Miscellaneous, Projetcs etc.
subject
keywords
PSP-CEC, UV, MALDI-TOF/MS
language
English
id
4699366
date added to LUP
2014-10-21 22:55:16
date last changed
2019-01-08 09:12:42
@misc{4699366,
  abstract     = {In this work, lipid nanoparticles were used as pseudostationary phase, PSP, for protein separation in capillary electrochromatography, CEC. The aim of the work was to use a new analytical separation technique for future analysis of protein drugs by using lipid nanoparticles as a pseudo-stationary phase with conventional UV-detection.
PSP-CEC was combined with UV-detection to study proteins elution. The tested proteins BSA, cytochrome c and insulin have been selected according to different hydrophobicities, sizes and isoelectric points (pI). A zwitter ionic buffer, MOPS, was used as an electrolyte at pH 7.4. The nanoparticles were tested for their compatibility with MALDI-TOF/MS (Matrix assisted laser desorption/ionization-time of flight). MADI-TOF/MS works with a suspension of lipid nanoparticles in MOPS that contains insulin and it can be a future possible detection method for PSP-CEC. All three proteins were successfully eluted with PSP-CEC combined with UV-detection. Nanoparticles coated with silver might be studied with “surface enhanced Raman spectroscopy”-detection.},
  author       = {Mannaa, Atef},
  keyword      = {PSP-CEC,UV,MALDI-TOF/MS},
  language     = {eng},
  note         = {Student Paper},
  title        = {Nanoparticle-based capillary electrochromatography},
  year         = {2013},
}