Peptidomics draws more and more attention in discovering useful biomarkers for early diagnosis of disease.However,there is lack of efficient quantification strategy in peptidome analysis.In this study,a strategy with ...Peptidomics draws more and more attention in discovering useful biomarkers for early diagnosis of disease.However,there is lack of efficient quantification strategy in peptidome analysis.In this study,a strategy with label-free quantification of the targeted endogenous peptides based on peak intensity usingμUPLC-Q-TOF-MS/MS was developed for quantitative peptidome analysis of human serum.Different amounts of standard BSA tryptic digesting peptides were added into the same serum extracts for evaluation of the developed strategy,and it was observed that the average relative error of the targeted peptides was 6.42%,which was superior to the result obtained directly by commercially available software PLGS.It was also demonstrated that this quantification strategy could obviously increase the detection sensitivity of the peptide by DDA analysis.Then,this strategy was applied to comparatively analyze the peptides extracted from the serum of HCC or breast cancer patients and healthy individuals,respectively.Peptides with charge states up to 5 and molecular weight over 4000 can be reliably identified and quantified.This quantitative analysis method based onμUPLC-Q-TOF-MS/MS exhibited superior sensitivity than that by MALDI-TOF-MS commonly used in peptidome analysis.Finally,some interesting endogenous peptides related to corresponding diseases were successfully obtained.展开更多
This review gives a broad glance on the progress of recent advances on proteolysis and peptide/protein separation by chroma-tographic strategies in the past ten years,covering the main research in these areas especial...This review gives a broad glance on the progress of recent advances on proteolysis and peptide/protein separation by chroma-tographic strategies in the past ten years,covering the main research in these areas especially in China.The reviewed research focused on enzymatic micro-reactors and peptide separation in bottom-up approaches,and protein and peptide separation in top-down approaches.The new enzymatic micro-reactor is able to accelerate proteolytic reaction rate from conventionally a couple of hours to a few seconds,and the multiple dimensional chromatographic-separation with various models or arrays could sufficiently separate the proteomic mixture.These advances have significantly promoted the research of protein/peptide separation and identification in proteomics.展开更多
基金support from the National Natural Science Foundation of China(Grant Nos.20735004&20975101)the State Key Basic Research Program of China(Grant Nos.2005CB522701&2007CB914102)+3 种基金the High Technology Research Pro-gram of China(Grant Nos.2006AA02A309&2008ZX10002-017)the Knowledge Innovation Program of Chinese Academy of Sciences(Grant Nos.KJCX2.YW.HO9&KSCX2-YW-R-079)the Knowledge Innova-tion Program of Dalian Institute of Chemical Physics to Zou HF and the China High Technology Research Program(Grant No.2008ZX1002-020)the National Natural Science Foundation of China(Grant Nos.20605022&90713017)to Ye ML
文摘Peptidomics draws more and more attention in discovering useful biomarkers for early diagnosis of disease.However,there is lack of efficient quantification strategy in peptidome analysis.In this study,a strategy with label-free quantification of the targeted endogenous peptides based on peak intensity usingμUPLC-Q-TOF-MS/MS was developed for quantitative peptidome analysis of human serum.Different amounts of standard BSA tryptic digesting peptides were added into the same serum extracts for evaluation of the developed strategy,and it was observed that the average relative error of the targeted peptides was 6.42%,which was superior to the result obtained directly by commercially available software PLGS.It was also demonstrated that this quantification strategy could obviously increase the detection sensitivity of the peptide by DDA analysis.Then,this strategy was applied to comparatively analyze the peptides extracted from the serum of HCC or breast cancer patients and healthy individuals,respectively.Peptides with charge states up to 5 and molecular weight over 4000 can be reliably identified and quantified.This quantitative analysis method based onμUPLC-Q-TOF-MS/MS exhibited superior sensitivity than that by MALDI-TOF-MS commonly used in peptidome analysis.Finally,some interesting endogenous peptides related to corresponding diseases were successfully obtained.
基金support from the Major State Basie Research Development Program(Grant No.2007CB914100)the National Natural Science Foundation of China(Grant Nos.20875016&20735005)Shanghai Projects(Grant Nos.08DZ2293601&B109)
文摘This review gives a broad glance on the progress of recent advances on proteolysis and peptide/protein separation by chroma-tographic strategies in the past ten years,covering the main research in these areas especially in China.The reviewed research focused on enzymatic micro-reactors and peptide separation in bottom-up approaches,and protein and peptide separation in top-down approaches.The new enzymatic micro-reactor is able to accelerate proteolytic reaction rate from conventionally a couple of hours to a few seconds,and the multiple dimensional chromatographic-separation with various models or arrays could sufficiently separate the proteomic mixture.These advances have significantly promoted the research of protein/peptide separation and identification in proteomics.
