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High-speed separation of proteins by microchip electrophoresis using a polyethylene glycol-coated plastic chip with a sodium dodecyl sulfate-linear polyacrylamide solution
Authors:Nagata Hideya  Tabuchi Mari  Hirano Ken  Baba Yoshinobu
Institution:Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, University of Tokushima, 21st Century COE Program, CREST(JST), Tokushima, Japan. hideya.nagata@aist.go.jp
Abstract:In this paper, we describe a method for size-based electrophoretic separation of sodium dodecyl sulfate (SDS)-protein complexes on a polymethyl methacrylate (PMMA) microchip, using a separation buffer solution containing SDS and linear polyacrylamide as a sieving matrix. We developed optimum conditions under which protein separations can be performed, using polyethylene glycol (PEG)-coated polymer microchips and electrokinetic sample injection. We studied the performance of protein separations on the PEG-coated PMMA microchip. The electrophoretic separation of proteins (21.5-116.0 kDa) was completed with separation lengths of 3 mm, achieved within 8 s on the PEG-coated microchip. This high-speed method may be applied to protein separations over a large range of molecular weight, making the PEG-coated microchip approach applicable to high-speed proteome analysis systems.
Keywords:Adsorption  Coating  Miniaturization  Polyethylene glycol  Polymethyl methacrylate microchip  Protein
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