Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Development of Ultra-High Pressure Capillary Reverse-Phase Liquid Chromatography/Tandem Mass Spectrometry for High-Sensitive and High-Throughput Proteomics

  • Kim, Min-Sik (Department of Chemistry and Center for Electro- & Photo-Responsive Molecules, Korea University) ;
  • Choie, Woo-Suk (Department of Chemistry and Center for Electro- & Photo-Responsive Molecules, Korea University) ;
  • Shin, Yong-Seung (Functional Proteomics Center, Korea Institute of Science and Technology, School of Life Sciences, Korea University) ;
  • Yu, Myeong-Hee (Functional Proteomics Center, Korea Institute of Science and Technology) ;
  • Lee, Sang-Won (Department of Chemistry and Center for Electro- & Photo-Responsive Molecules, Korea University)
  • Published : 2004.12.20
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Abstract

Recently mass spectrometry and separation methods such as liquid chromatography have become major tools in the field of proteomics. In this report, we describe in detail our efforts to develop ultra-high pressure capillary reverse-phase liquid chromatography (cRPLC) and its online coupling to a mass spectrometer by a nanoelectrospray (nanoESI) interface. The RPLC system is constructed in house to deliver LC solvents at the pressure up to 20,000 psig, which is four times higher than conventional RPLC systems. The high operation pressure allows the efficient use of packed micro-capillary columns (50, 75 and 150 ${\mu}$m i.d., up to 1.5 m long). We will discuss the effect of column diameter on the sensitivity of cRPLC/MS/MS experiments and the utility of the developed technique for proteome analysis by its application in the analysis of proteome samples having different levels of complexity.

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References

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