Ultra-fast two-dimensional microchip electrophoresis using SDS micro-CGE and microemulsion electrokinetic chromatography for protein separations.
A poly(methyl methacrylate) microfluidic chip was used to perform a two-dimensional (2-D) separation of a complex protein mixture in short development times. The separation was performed by combining sodium dodecyl sulfate micro-capillary gel electrophoresis (SDS micro-CGE) with microemulsion electrokinetic chromatography (micro-MEEKC), which were used for the first and second dimensions, respectively. Fluorescently labeled Escherichia coli cytosolic proteins were profiled by this 2-D approach with the results compared to a similar 2-D separation using SDS micro-CGE x micro-MEKC (micelle electrokinetic chromatography). The relatively short column lengths (effective length = 10 mm) for both dimensions were used to achieve separations requiring only 220 s of development time. High spot production rates (131 +/- 11 spots min(-1)) and reasonable peak capacities (481 +/- 18) were generated despite the fact that short columns were used. In addition, the use of mu-MEEKC in the second dimension was found to produce higher peak capacities compared to micro-MEKC (481 +/- 18 for micro-MEEKC and 332 +/- 17 for micro-MEKC) due to the higher plate numbers associated with micro-MEEKC.
Osiri
John K
JK
Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA.
Shadpour
Hamed
H
Soper
Steven A
SA
D001426:Bacterial Proteins / Q000302:isolation & purification*; D020374:Chromatography, Micellar Electrokinetic Capillary; D019075:Electrophoresis, Capillary; D015180:Electrophoresis, Gel, Two-Dimensional; D046209:Electrophoresis, Microchip; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism; D008823:Micelles; D019904:Polymethyl Methacrylate / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*
D001426:Bacterial Proteins; D008823:Micelles; D012967:Sodium Dodecyl Sulfate; D019904:Polymethyl Methacrylate