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Miniaturized two‐dimensional capillary electrophoresis on a microchip for analysis of the tryptic digest of proteins
Author(s) -
Cong Yongzheng,
Zhang Lihua,
Tao Dingyin,
Liang Yu,
Zhang Weibing,
Zhang Yukui
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700444
Subject(s) - isoelectric focusing , capillary electrophoresis , chromatography , chemistry , microchannel , electrophoresis , analytical chemistry (journal) , resolution (logic) , capillary action , materials science , nanotechnology , computer science , biochemistry , artificial intelligence , composite material , enzyme
A two‐dimensional capillary electrophoresis platform, combining isoelectric focusing (IEF) and capillary zone electrophoresis (CZE), was established on a microchip with the channel width and depth as 100 μm and 40 μm, respectively. With polyacrylamide as permanent coating, EOF in the microchannel, which could impair the separation, was decreased to 3.4×10 –9 m 2 ·V –1 ·s –1 , about 1/10 of that obtained in the uncoated set‐up. During the separation, peptides were first focused by IEF in the first dimensional channel, and then directly driven into the perpendicular channel by controlling the applied voltages, and separated by CZE. Effects of various experimental parameters, including the electric field strength, channel length, and injection frequency from the first to the second dimensional separation channel, were studied. Under optimized condition, the digests of BSA and proteins extracted from E. coli were separated, and a peak capacity of 540 was obtained, which was far greater than that obtained by each single dimensional separation. All these results showed the promise of multidimensional separation on a microchip for the high‐throughput and high‐resolution analysis of complex samples.