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KMID : 0370220100540050377
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Yakhak Hoeji
2010 Volume.54 No. 5 p.377 ~ p.386
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Comparison of 2-D RP-RP MS/MS with 1-D RP MS/MS for Proteomic Analysis
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Moon Pyoung-Gon
Cho Young-Eun
Baek Moon-Chang
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Abstract
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Single-dimensional (1-D) and two-dimensional (2-D) LC methods were utilized to separate peptides from various sources followed by MS/MS analysis. Two-dimensional ultra-high performance liquid chromatography is a useful tool for proteome analysis, providing a greater peak capacity than 1-D LC. The most popular 2-D LC approach used today for proteomic research combines strong cation exchange and reversed-phase LC. We have evaluated an alternative mode for 2-D LC of peptides using 2-D RP-RP nano UPLC Q-TOF Mass Spectrometry, employing reversed-phase columns in both separation dimensions. As control experiments, we identified 129 proteins in 1-D LC and 322 proteins in 2-D LC from E. coli extract peptides. Furthermore, we applied this method to rat primary hepatocyte and a total of 170 proteins were identified from 1-D LC, and 527 proteins were identified from all 2-D LC system. The in-depth protein profiling established by this 2-D LC MS/MS from rat primary hepatocyte could be a very useful reference for future applications in regards to drug induced liver toxicity.
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KEYWORD
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2-D LC MS/MS, RP-RP, hepatocyte
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