High-speed separation of proteins by microchip electrophoresis using a polyethylene glycol-coated plastic chip with a sodium dodecyl sulfate-linear polyacrylamide solution.
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.
Nagata
Hideya
H
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
Tabuchi
Mari
M
Hirano
Ken
K
Baba
Yoshinobu
Y
D000180:Acrylic Resins / Q000737:chemistry*; D046209:Electrophoresis, Microchip / Q000379:methods*; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D019904:Polymethyl Methacrylate / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D000099166:Polyacrylamides
D000180:Acrylic Resins; D011092:Polyethylene Glycols; D019904:Polymethyl Methacrylate; D011506:Proteins; D012967:Sodium Dodecyl Sulfate; D012996:Solutions; D000099166:Polyacrylamides