title: Identification of low-abundance proteins via fractionation of the urine proteome with weak anion exchange chromatography.
abstract: BACKGROUND
  Low-abundance proteins are difficultly observed on the two-dimensional gel electrophoresis (2-DE) maps of urine proteome, because they are usually obscured by high-abundance proteins such as albumin and immunoglobulin. In this study, a novel fractionation method was developed for enriching low-abundance proteins by removing high-abundance proteins and progressive elution with salts of various concentrations.
  
  
  RESULTS
  Stepwise weak anion exchange (WAX) chromatography, which applied DEAE-Sephacel resin with non-fixed volume elution, was used to fractionate urine proteome prior to performing 2-DE. Urine proteome was separated into four fractions by progressively eluting the column with 0 M, 50 mM, 100 mM, and 1 M NaCl solutions. Most of the heavy and light immunoglobulin chains appeared in the eluent. After the high-abundance proteins were removed, various low-abundance proteins were enriched and could be easily identified. The potential of this method for obtaining diversified fractionations was demonstrated by eluting the column separately with Na2SO4 and MgCl2 solutions. The 2-DE maps of the fractions eluted with these different salt solutions of identical ionic strength revealed markedly different stain patterns.
  
  
  CONCLUSIONS
  The present study demonstrated that this fractionation method could be applied for purposes of enriching low-abundance proteins and obtaining diversified fractionations of urine, and potentially other proteomes.
authors.affiliation: Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.|Department of Urology, Buddhist Da Lin Tzu Chi General Hospital, Chiayi, Taiwan.
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Lu
authors.affiliation: Department of Beauty Science, Meiho University, Pingtung, Taiwan.
authors.forename: Yu-Jen
authors.initials: YJ
authors.lastname: Wu
authors.affiliation: Department of Beauty Science, Meiho University, Pingtung, Taiwan.
authors.forename: Cheng-Chi
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Graduate Institute of Biotechnology, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Jue-Liang
authors.initials: JL
authors.lastname: Hsu
authors.affiliation: Department of Food Science and Nutrition, Meiho University, Pingtung, Taiwan.
authors.forename: Jiing-Chuan
authors.initials: JC
authors.lastname: Chen
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Jeff Yi-Fu
authors.initials: JY
authors.lastname: Chen
authors.affiliation: Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.|Department of Urology, Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Chun-Hsiung
authors.initials: CH
authors.lastname: Huang
authors.affiliation: Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.|Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Ying-Chin
authors.initials: YC
authors.lastname: Ko