Microchip electrophoretic protein separation using electroosmotic flow induced by dynamic sodium dodecyl sulfate-coating of uncoated plastic chips.
Separation of sodium dodecyl sulfate (SDS)-protein complexes is difficult on plastic microchips due to protein adsorption onto the wall. In this paper, we elucidated the reasons for the difficulties in separating SDS-protein complexes on plastic microchips, and we then demonstrated an effective method for separating proteins using polymethyl methacrylate (PMMA) microchips. Separation difficulties were found to be dependent on adsorption of SDS onto the hydrophobic surface of the channel, by which cathodic electroosmotic flow (EOF; reversed flow) was generated. Our developed method effectively utilized the reversed flow from this cathodic EOF as a driving force for sample proteins using permanently uncoated but dynamic SDS-coated PMMA microchips. High-speed (6 s) separation of proteins and peptides up to 116 kDa was successfully achieved using this system.
Nagata
Hideya
H
Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, The University of Tokushima, The 21ST Century COE Program, CREST(JST), Tokushima, Japan. hideya.nagata@aist.go.jp
Tabuchi
Mari
M
Hirano
Ken
K
Baba
Yoshinobu
Y
D000327:Adsorption; D020099:Coated Materials, Biocompatible; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods; D057927:Hydrophobic and Hydrophilic Interactions; D010969:Plastics; D019904:Polymethyl Methacrylate; D011506:Proteins / Q000302:isolation & purification*; D012212:Rheology; D012967:Sodium Dodecyl Sulfate
D020099:Coated Materials, Biocompatible; D010969:Plastics; D011506:Proteins; D012967:Sodium Dodecyl Sulfate; D019904:Polymethyl Methacrylate