A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography system for high-throughput proteomics analysis

Hangyeore Lee; Jung Hwa Lee; Hokeun Kim; Su Jin Kim; Jingi Bae; Hark Kyun Kim; Sang Won Lee

doi:10.1016/j.chroma.2013.12.084

A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography system for high-throughput proteomics analysis

Hangyeore Lee, Jung Hwa Lee, Hokeun Kim, Su Jin Kim, Jingi Bae, Hark Kyun Kim, Sang Won Lee

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	83-89
Number of pages	7
Journal	Journal of Chromatography A
Volume	1329
DOIs	https://doi.org/10.1016/j.chroma.2013.12.084
Publication status	Published - 2014 Feb 14

Bibliographical note

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 ).

Keywords

Bottom-up proteomics
Dual-online LC
Multidimensional separation
Tandem mass spectrometry

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Organic Chemistry

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10.1016/j.chroma.2013.12.084

Cite this

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title = "A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography system for high-throughput proteomics analysis",

abstract = "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.",

keywords = "Bottom-up proteomics, Dual-online LC, Multidimensional separation, Tandem mass spectrometry",

author = "Hangyeore Lee and Lee, {Jung Hwa} and Hokeun Kim and Kim, {Su Jin} and Jingi Bae and Kim, {Hark Kyun} and Lee, {Sang Won}",

note = "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 ). ",

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doi = "10.1016/j.chroma.2013.12.084",

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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 ).

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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.

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A fully automated dual-online multifunctional ultrahigh pressure liquid chromatography system for high-throughput proteomics analysis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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