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Novel negatively charged tentacle-type polymer coating for on-line preconcentration of proteins in CE.

Authors :
Xu L
Dong XY
Sun Y
Source :
Electrophoresis [Electrophoresis] 2009 Feb; Vol. 30 (4), pp. 689-95.
Publication Year :
2009

Abstract

A novel negatively charged tentacle-type polymer-coated capillary column was fabricated and applied for on-line extraction and preconcentration of proteins. The polymer coating was prepared by glycidyl-methacrylate graft polymerization in a silanized capillary column and the following sulfonic acid group functionalization. It had high surface area and offered high phase ratio for protein adsorption. In addition, the polymer-coated capillary column provided more stable EOF than a bare uncoated capillary. These features of the polymer coating facilitated the extraction of proteins through electrostatic interactions. This was used to extract proteins. The extracted analytes were then desorbed and focused by EOF in the direction opposite to the sample injection flow for subsequent CE. With this procedure, over 1500-fold sensitivity enhancement was realized for myoglobin (MB) as compared with a normal capillary zone electrophoresis. By comparison of the peak areas of the enriched protein, it was found that the polymer-coated column could capture proteins about 30 times more than the uncoated column. In addition, the separation of a protein mixture containing 0.4 microg/mL of MB and 0.4 microg/mL of insulin was demonstrated by the on-line preconcentration and electrophoretic separation with the polymer-coated column.

Details

Language :
English
ISSN :
1522-2683
Volume :
30
Issue :
4
Database :
MEDLINE
Journal :
Electrophoresis
Publication Type :
Academic Journal
Accession number :
19170054
Full Text :
https://doi.org/10.1002/elps.200800558