title: An individual urinary proteome analysis in normal human beings to define the minimal sample number to represent the normal urinary proteome.
abstract: BACKGROUND
  The 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.
  
  
  METHODS
  In 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.
  
  
  RESULTS
  For 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.
  
  
  CONCLUSIONS
  For 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.
authors.affiliation: Department of Nephrology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, No. 1 Shuaifuyan, Wangfujing Street, Beijing, China.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Physiology and Pathophysiology, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences/School of Basic Medicine Peking Union Medical College, 5 Dong Dan San Tiao, Beijing, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Shao
authors.affiliation: Clinical laboratory, China-Japan Friendship Hospital, 2 Yinghua Dongjie, Hepingli, Beijing, China.
authors.forename: Lilong
authors.initials: L
authors.lastname: Wei
authors.affiliation: Core facility of instrument, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences/School of Basic Medicine Peking Union Medical College, 5 Dong Dan San Tiao, Beijing, China.
authors.forename: Jindan
authors.initials: J
authors.lastname: Duan
authors.affiliation: Core facility of instrument, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences/School of Basic Medicine Peking Union Medical College, 5 Dong Dan San Tiao, Beijing, China.
authors.forename: Shuzhen
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of Nephrology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, No. 1 Shuaifuyan, Wangfujing Street, Beijing, China.
authors.forename: Xuewang
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Nephrology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, No. 1 Shuaifuyan, Wangfujing Street, Beijing, China.
authors.forename: Mingxi
authors.initials: M
authors.lastname: Li
authors.affiliation: Core facility of instrument, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences/School of Basic Medicine Peking Union Medical College, 5 Dong Dan San Tiao, Beijing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun