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Protein and peptide separations on high surface area capillaries
Author(s) -
Pesek Joseph J.,
Matyska Maria T.,
Swedberg Sally,
Udivar Sharmila
Publication year - 1999
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/(sici)1522-2683(19990801)20:12<2343::aid-elps2343>3.0.co;2-9
Subject(s) - capillary electrochromatography , capillary action , capillary electrophoresis , chromatography , chemistry , resolution (logic) , analytical chemistry (journal) , electrophoresis , electrochromatography , materials science , composite material , artificial intelligence , computer science
A 50 μm capillary that has been etched with ammonium hydrogen difluoride is evaluated as a separation medium for capillary electrochromatography. For a tryptic digest of transferrin, the etched capillary gave better resolution (more peaks in the overall peptide map) and longer retention than separations done under identical experimental conditions on an unetched fused‐silica capillary. Resolution on the etched capillary was improved by lowering the voltage from 300 to 267 V/cm. A four‐component protein sample also resulted in longer retention on an echted capillary than on an unetched fused‐silica capillary that were both coated with Polybrene. After correction for differences in electroosmotic flow between the two capillaries, the calculated electrophoretic mobilities for all four proteins were lower on the etched capillary than on the unetched fused‐silica capillary. The results of both the tryptic digest and protein experiments strongly indicate the presence of chromatographic effects on the etched capillary that contribute to the increased retention and improved resolution with respect to the unetched fused‐silica capillary.