CHROMATOGRAPHY
CHROMATOGRAPHY, Vol. 26 (2005), No. 1, pp. 23-28
Original
Automatic Protein Separation by Microchip Electrophoresis Using Quartz Chip
Hideya Nagata1*, Mari Tabuchi1, Ken Hirano1 and Yoshinobu Baba1,2,3
1Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, The University of Tokushima, The 21st Century COE Program, CREST(JST), Tokushima, Japan
2Single.Molecule Bioanalysis Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Japan 3Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Nagoya, Japan
Abstract:
MCE using a quartz microchip and an LIF detector was applied to the analysis of proteins. In this study, the experimental conditions were optimized to achieve faster separation and higher resolution of SDS.denatured proteins chemically labeled with fluorescent dye. Proteins (MW 5.7.116. 0 kDa) were completely separated (Rs=1.1.4.4) within 160 s under the optimized conditions using 4.4% linear polyacrylamide at 100 V/cm and the plate number was 0.1.1.4×106 plates/m. Increasing the separation voltage decreased analysis time (70.90 s at 200.240 V/cm) but decreased the resolution relative to the optimal conditions. The detection limit achieved (0.13 ng/μL) is almost 150 times lower than that of the conventional MCE system.
Keywords: Protein / Microchip Electrophoresis / Quartz microchip / Linear polyacrylamide / Sequential analysis