An individual urinary proteome analysis in normal human beings to define the minimal sample number to represent the normal urinary proteome

Abstract

BackgroundThe urinary proteome has been widely used for biomarker discovery. A urinary proteome database from normal humans can provide a background for discovery proteomics and candidate proteins/peptides for targeted proteomics. Therefore, it is necessary to define the minimum number of individuals required for sampling to represent the normal urinary proteome.MethodsIn this study, inter-individual and inter-gender variations of urinary proteome were taken into consideration to achieve a representative database. An individual analysis was performed on overnight urine samples from 20 normal volunteers (10 males and 10 females) by 1DLC/MS/MS. To obtain a representative result of each sample, a replicate 1DLCMS/MS analysis was performed. The minimal sample number was estimated by statistical analysis.ResultsFor qualitative analysis, less than 5% of new proteins/peptides were identified in a male/female normal group by adding a new sample when the sample number exceeded nine. In addition, in a normal group, the percentage of newly identified proteins/peptides was less than 5% upon adding a new sample when the sample number reached 10. Furthermore, a statistical analysis indicated that urinary proteomes from normal males and females showed different patterns. For quantitative analysis, the variation of protein abundance was defined by spectrum count and western blotting methods. And then the minimal sample number for quantitative proteomic analysis was identified.ConclusionsFor qualitative analysis, when considering the inter-individual and inter-gender variations, the minimum sample number is 10 and requires a balanced number of males and females in order to obtain a representative normal human urinary proteome. For quantitative analysis, the minimal sample number is much greater than that for qualitative analysis and depends on the experimental methods used for quantification.

Identification of a Three-Biomarker Panel in Urine for Early Detection of Pancreatic Adenocarcinoma

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A Proteomic Analysis of Individual and Gender Variations in Normal Human Urine and Cerebrospinal Fluid Using iTRAQ Quantification

“…Urine and CSF are two important biofluids. Indeed, urinary proteome contains useful information about kidneys, urinary tract and other organs [ 1 , 2 ], and urinary proteome has been used to identify biomarkers of bladder cancer [ 3 ], chronic kidney disease [ 4 ], and diabetic nephropathy [ 5 ]. CSF can provide information about central nervous system (CNS), and this fluid has been used for the analysis of a number of CNS diseases, including Alzheimer’s disease (AD) [ 6 , 7 ], Parkinson’s disease [ 8 ], and pediatric medulloblastoma [ 9 ].…”

Urine sample preparation for proteomic analysis

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