Fully automated multifunctional ultrahigh pressure liquid chromatography system for advanced proteome analyses.
A multifunctional liquid chromatography system that performs 1-dimensional, 2-dimensional (strong cation exchange/reverse phase liquid chromatography or SCX/RPLC) separations and online phosphopeptide enrichment using a single binary nanoflow pump has been developed. With a simple operation of a function selection valve equipped with a SCX column and a TiO2 (titanium dioxide) column, a fully automated selection of three different experiment modes was achieved. Because the current system uses essentially the same solvent flow paths, the same trap column, and the same separation column for reverse-phase separation of 1D, 2D, and online phosphopeptides enrichment experiments, the elution time information obtained from these experiments is in excellent agreement, which facilitates correlating peptide information from different experiments. The final reverse-phase separation of the three experiments is completely decoupled from all of the function selection processes; thereby salts or acids from SCX or TiO2 column do not affect the efficiency of the reverse-phase separation.
Department of Chemistry, Research Institute for Natural Sciences, Korea University, Seoul 136-701, South Korea.
Jung Hwa
JH
Lee
Seok-Won
SW
Hyung
Dong-Gi
DG
Mun
Hee-Jung
HJ
Jung
Hokeun
H
Kim
Hangyeore
H
Lee
Su-Jin
SJ
Kim
Kyong Soo
KS
Park
Ronald J
RJ
Moore
Richard D
RD
Smith
Sang-Won
SW
Lee
D000595:Amino Acid Sequence; D057205:Automation, Laboratory / Q000295:instrumentation*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D002852:Chromatography, Ion Exchange / Q000295:instrumentation; D056148:Chromatography, Reverse-Phase; D006801:Humans; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification*; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification*; D052616:Solid Phase Extraction / Q000295:instrumentation; D053719:Tandem Mass Spectrometry / Q000295:instrumentation*
D010750:Phosphoproteins; D020543:Proteome