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Whole‐column imaging capillary electrophoresis of proteins with a short capillary
Author(s) -
Wu XingZheng,
Pawliszyn Janusz
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200202)23:4<542::aid-elps542>3.0.co;2-b
Subject(s) - capillary action , capillary electrophoresis , capillary electrochromatography , chromatography , chemistry , analytical chemistry (journal) , analyte , sodium dodecyl sulfate , isoelectric focusing , materials science , composite material , biochemistry , enzyme
Whole‐column imaging capillary electrophoresis with a short capillary is discussed. A short capillary (3–6 cm) coated with either fluorocarbon or polyacrylamide was used as a separation capillary. The whole capillary was illuminated with 280 nm light, and the transmitted light was monitored by a linear charge‐coupled device (CCD). For the short capillary, hydrodynamic flow caused by a subtle height difference between the anodic and cathodic reservoirs affected the sample migration in the capillary greatly. Several sample injection methods, including use of a cross connection, sealing of the capillary ends with a gel, and use of a gel‐filled capillary, have been discussed. The experimental results showed that the peak height decreased and peak width increased with the electromigration distance. Therefore, higher sensitivity was obtained in a short capillary rather than a long capillary. The whole‐column imaging CE with the short capillary has been applied for the study of conjugation reactions of protein cytochrome c with sodium dodecyl sulfate (SDS) and the dye Congo Red. The method has also been used for in situ monitoring of the electrophoretic protein desorption process. Our technique is a unique tool for the study of protein binding reactions and the interaction between analyte and inner wall of the capillary.