Optimization Of C18 Solid Phase Extraction Columns And Analysis Of Differential Proteomics Of Urine In Coronary Heart Disease

Objective:To develop a timesaving and easy operating Reverse Phase (RP) chromatography method.Methods:We adopted Thermo Pierce(?) RP C18 Tip to separate small amount hippocampus peptide mixtures and to compare with high performance liquid chromatography (HPLC). According to the separation performance of 4 ACN gradient optimization methods, we determined the best ACN concentration gradient.Results:The results showed that, the experiment took only 10 min by separating with eight ACN concentration gradient, which accounted 1/4 for HPLC. But as for the identified proteins, RP C18 Tip accounted 85.5% for HPLC. ACN gradient of 5,15,20 and 90% had best protein repeatability (70%) and result for separating 30 μg peptides.Conclusion:This method is easy to operate, timesaving and has low cost. It could be used into pretreatment of small amount complex proteomic samples.Objective:Urinary proteome have been caught much attention due to non-invasive sampling and no special preparation of urine. Previous work based on capillary electrophoresis-coupled mass spectrometry (CE-MS) has provided useful urinary proteome data of atherosclerosis on peptide level. We tried to present a comprehensive proteomic analysis on protein level.Methods:iTRAQ-based 2D LC-MS/MS was used to analyze the urinary proteome from atherosclerotic patients and controls.Results:Total 856 urinary proteins were identified and 737 proteins were quantified.90 differential proteins were found between the two groups (fold change,>1.5). Functional analysis revealed that lipid metabolism, infection and inflammatory response and glycometabolism were highlighted.Conclusion:Urine could reflect both intracellular and extracellular changes of atherosclerosis, which might be a good source for disease biomarkers.