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芯片修饰3
462 Y. Cong et al. J. Sep. Sci. 2009, 32, 462 – 465
Yongzheng Cong1, 2 Short Communication
Yu Liang1
Lihua Zhang1
Weibing Zhang1 Improved protein separation by microchip
Yukui Zhang1 isoelectric focusing with stepwise gradient of
1National Chromatographic R. electric field strength
and A. Center, Key Lab of
Separation Science for Analytical
A stepwise gradient of electric field strength was proposed for microchip IEF-based
Chemistry, Dalian Institute of
Chemical Physics, Chinese protein separation, by which after focusing at low voltages, IEF was performed by
Academy of Sciences, Dalian, applying higher separation voltages step-by-step. A linear relationship between the
China focusing time and the inverse of the electric field strength was found. In addition,
2Graduate School of Chinese the conductivity of an established pH gradient showed a negative but nonlinear cor-
Academy of Sciences, Beijing, relation with the applied voltage. Based on the above-mentioned results, a stepwise
China
gradient of electric field strength, ranging from 160 to 1500 V/cm was applied in
the separation of proteins extracted from Escherichia coli in a straight glass microchip
channel permanently coated by polyacrylamide. Compared to the conventional sep-
aration performed under a constant field strength of 750 V/cm, the increased step-
wise gradient of electric field strength resulted in improved resolution and
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