Novel Separation and Identification Techniques for Proteome Study from Qualitative to Quantitative Analysis

In the past decade, proteomics has been seen rapid advance development due to the emerging of new analytical methods. To date, besides qualitative analysis, protein quantification, the driving force of proteomics, has been widely applied to reveal biological processes. At present, the hyphenation of isotopic labeling and mass spectrometry (MS) techniques is the dominant approach in quantitative proteomics. Many isotopic reagents were developed and applied in quantitative proteomics, such as ICAT, SILAC and iTRAQ, etc. If internal standards were added into the sample, absolute quantification information can be obtained, for example, AQUA, QconCAT etc. Besides these methods, label free and ICP-MS methods have also been commonly used for protein quantification. However, the present proteome quantitative techniques suffer from low sensitivity, low accuracy, and low throughput. Therefore, in our recent work, much effort has been made to solve these problems, by the development of new sample preparation materials with high recovery, separation matrices with high efficiency, integrated platforms with good reproducibility and high throughput, and labeling strategies with high accuracy, by which the accuracy for proteome quantitative analysis could be obviously improved.In the past decade, proteomics has been seen rapid advance development due to the emerging of new analytical methods. To date, besides qualitative analysis, protein quantification, the driving force of proteomics, has been widely applied to reveal biological processes. At present, the hyphenation of isotopic labeling and mass spectrometry (MS) techniques is the dominant approach in quantitative proteomics. Many isotopic reagents were developed and applied in quantitative proteomics, such as ICAT, SILAC and iTRAQ, etc. If internal standards were added into the sample, absolute quantification information can be obtained, for example, AQUA, QconCAT etc. Besides these methods, label free and ICP-MS methods have also been commonly used for protein quantification. However, the present proteome quantitative techniques suffer from low sensitivity, low accuracy, and low throughput. Therefore, in our recent work, much effort has been made to solve these problems, by the development of new sample preparation materials with high recovery, separation matrices with high efficiency, integrated platforms with good reproducibility and high throughput, and labeling strategies with high accuracy, by which the accuracy for proteome quantitative analysis could be obviously improved