A microfabricated fluidic device for performing two-dimensional liquid-phase separations.
A microfabricated fluidic device that combines micellar electrokinetic chromatography and high-speed open-channel electrophoresis on a single structure for the rapid automated two-dimensional analysis of peptides has been devised and demonstrated. The microchip operates by rapidly sampling and analyzing effluent in the second dimension from the first dimension. Second-dimension analyses are performed and completed every few seconds, with total analysis times of less than 10 min for tryptic peptides. The peak capacity of the two-dimensional separations has been estimated to be in the 500-1000 range. The orthogonality of the separation techniques, an important factor for maximizing peak capacity or resolution elements, was verified by examining each technique independently for peptide separations. The two-dimensional separation strategy was found to greatly increase the resolving power over that obtained for either dimension alone.
Rocklin
R D
RD
Chemical and Analytical Sciences Division, Oak Ridge National Laboratory, Tennessee 37831-6142, USA.
Ramsey
R S
RS
Ramsey
J M
JM
D002845:Chromatography; D019075:Electrophoresis, Capillary; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D008904:Miniaturization; D010446:Peptide Fragments; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*
D005456:Fluorescent Dyes; D010446:Peptide Fragments; D010455:Peptides; D011506:Proteins; D016650:Fluorescein-5-isothiocyanate