Multidimensional Protein Profiling Technology and Its Application to Human Plasma Proteome

• DOI: 10.1021/pr030007q
• Publication Date: 2004-08-09T04:16:25Z
• Authors: Kiyonaga Fujii; Tomoyo Nakano; Takeshi Kawamura; Fumihiko Usui; Yasuhiko Bando; Rong Wang; Toshihide Nishimura
• Keywords: Proteome

In clinical and diagnostic proteomics, it is essential to develop a comprehensive robust system for proteome analysis. Although multidimensional liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems have been recently developed as powerful tools especially identification of protein complexes, these still some drawbacks in their application research that requires an analysis large number human samples. Therefore, this study, we constructed technically simple high throughput profiling comprising two-dimensional (2D)-LC/MS/MS which integrates both strong cation exchange (SCX) chromatography μLC/MS/MS with micro-flowing reversed-phase chromatography. Using the second dimensional chromatography, SCX separation has optimized off-line first peptide fractionation. To evaluate performance 2D-LC/MS/MS system, results detection proteins were compared using digests mixtures 6 authentic those obtained one-dimensional system. The fragments detected coverage sequence found be more than double through use our newly built Furthermore, applied plasma order examine its feasibility proteomics. experimental revealed 174 from one serum sample depleted HSA IgG corresponds only 1 μL plasma, total run time was less half day, indicating fairly possibility practicing proteomics manner. Keywords: •