TY - JOUR
T1 - A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography system for high-throughput proteomics analysis
AU - Lee, Hangyeore
AU - Lee, Jung Hwa
AU - Kim, Hokeun
AU - Kim, Su Jin
AU - Bae, Jingi
AU - Kim, Hark Kyun
AU - Lee, Sang Won
N1 - Funding Information:
This work was supported in part by the Proteogenomic Research Program through the National Research Foundation of Korea funded by the Korean Ministry of Science , ICT & Future Planning, the Converging Research Center Program through the Ministry of Science , ICT and Future Planning, Korea ( 2013K000443 ). This work was also funded by a grant from the National Project for Personalized Genomic Medicine, Ministry for Health & Welfare, Republic of Korea ( A111218-CP02 ). We also acknowledge the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF20100020209 ).
PY - 2014/2/14
Y1 - 2014/2/14
N2 - A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography (DO-MULTI-UPLC) system has been developed for high throughput proteome analyses of complex peptide mixtures. The system employs two online solid phase extraction (SPE) columns (150. μm inner diameter. ×. 3. cm), two capillary reverse phase (RP) columns (75. μm. ×. 100. cm) and a strong cation exchange (SCX) column (150. μm. ×. 15. cm) on a single system utilizing one binary pump and one isocratic pump. With the automated operation of six switching valves, the selection of LC experiments between single-dimensional RPLC and online two-dimensional SCX/RPLC were achieved automatically, without manual intervention, while two RPLC columns were used independently and alternatively. By essentially removing the dead time for column equilibration between experiments, in either 1D mode or 2D experimental mode, the current system was demonstrated to increase the experimental throughput by about two folds, while keeping the inter-column reproducibility of peptide elution time in less than 1% of gradient time. The advantageous features of the proposed system were demonstrated by its application to proteome samples of varying complexities.
AB - A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography (DO-MULTI-UPLC) system has been developed for high throughput proteome analyses of complex peptide mixtures. The system employs two online solid phase extraction (SPE) columns (150. μm inner diameter. ×. 3. cm), two capillary reverse phase (RP) columns (75. μm. ×. 100. cm) and a strong cation exchange (SCX) column (150. μm. ×. 15. cm) on a single system utilizing one binary pump and one isocratic pump. With the automated operation of six switching valves, the selection of LC experiments between single-dimensional RPLC and online two-dimensional SCX/RPLC were achieved automatically, without manual intervention, while two RPLC columns were used independently and alternatively. By essentially removing the dead time for column equilibration between experiments, in either 1D mode or 2D experimental mode, the current system was demonstrated to increase the experimental throughput by about two folds, while keeping the inter-column reproducibility of peptide elution time in less than 1% of gradient time. The advantageous features of the proposed system were demonstrated by its application to proteome samples of varying complexities.
KW - Bottom-up proteomics
KW - Dual-online LC
KW - Multidimensional separation
KW - Tandem mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=84895107234&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2013.12.084
DO - 10.1016/j.chroma.2013.12.084
M3 - Article
C2 - 24412000
AN - SCOPUS:84895107234
SN - 0021-9673
VL - 1329
SP - 83
EP - 89
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -