Body fluid proteome is the most informative proteome from a medical viewpoint. But the lack of accurate quantitation method for complicated body fluid limited its application in disease research and biomarker discovery. To address this problem, we introduced a novel strategy, in which SILAC-labeled mouse serum was used as internal standard for human serum and urine proteome analysis. The SILAC-labeled mouse serum was mixed with human serum and urine, and multidimensional separation coupled with tandem mass spectrometry (IEF-LC-MS/MS) analysis was performed. The shared peptides between two species were quantified by their SILAC pairs, and the human-only peptides were quantified by mouse peptides with coelution. The comparison for the results from two replicate experiments indicated the high repeatability of our strategy. Then the urine from Immunoglobulin A nephropathy patients treated and untreated was compared by this quantitation strategy. Fifty-three peptides were found to be significantly changed between two groups, including both known diagnostic markers for IgAN and novel candidates, such as Complement C3, Albumin, VDBP, ApoA, 1 and IGFBP7. In conclusion, we have developed a practical and accurate quantitation strategy for comparison of complicated human body fluid proteome. The results from such strategy could provide potential disease-related biomarkers for evaluation of treatment.
[1]Chinese Acad Sci, Inst Biochem & Cell Biol, Key Lab Syst Biol, Shanghai Inst Biol Sci, Shanghai 200031, Peoples R China.
[2]Shanghai Univ Tradit Chinese Med, Longhua Hosp, Dept Nephrol, Shanghai 200032, Peoples R China.
[3]Univ Michigan, Med Ctr, Dept Internal Med, Ctr Cardiovasc, Ann Arbor, MI 48109 USA.
(8.0+极速模式)