The proteomic analysis improved by cleavage kinetics‐based fractionation of tryptic peptides |
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| Authors: | Yanbo Pan Jiawei Mao Zhenzhen Deng Mingming Dong Yangyang Bian Hanfa Zou |
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| Affiliation: | 1. Key Lab of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China;2. University of Chinese Academy of Sciences, Beijing, P. R. China |
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| Abstract: | Selective enrichment of specific peptides is an effective way to identify low abundance proteins. Fractionation of peptides prior to mass spectrometry is another widely used approach to reduce sample complexity in order to improve proteome coverage.In this study, we designed a multi‐stage digestion strategy to generate peptides with different trypsin cleavage kinetics. It was found that each of the collected peptide fractions yielded many new protein identifications compared to the control group due to the reduced complexity. The overlapping peptides identified between adjacent fractions were very low, indicating that each fraction had different sets of peptides. The multi‐stage digestion strategy separates tryptic peptides with different cleavage kinetics while RPLC separates peptides with different hydrophobicity. These two separation strategies were highly orthogonal, and showed an effective multidimensional separation to improve proteome coverage. |
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| Keywords: | Fractionation of peptides Mass spectrometry Proteomic analysis Technology Trypsin digestion kinetics |
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