A method for isolation and identification of urinary biomarkers in patients with diabetic nephropathy

Wayne G. Fisher, Jessica E. Lucas, Uzma F. Mehdi, Danna W. Qunibi, Harold R. Garner, Kevin P. Rosenblatt, Robert D. Toto

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Purpose: The poor performance of current tests for predicting the onset, progression and treatment response of diabetic nephropathy has engendered a search for more sensitive and specific urinary biomarkers. Our goal was to develop a new method for protein biomarker discovery in urine from these patients. Experimental design: We analyzed urine from normal subjects and patients with early and advanced nephropathy. Proteins were separated using a novel analysis process including immunodepletion of high-abundance proteins followed by two-stage LC fractionation of low-abundance proteins. The proteins in the fractions were sequenced using MS/MS. Results: Immunodepletion of selected high-abundance proteins followed by two-stage LC produced approximately 700 fractions, each less complex and more amenable to analysis than the mixture and requiring minimal processing for MS identification. Comparison of fractions between normal and diabetic nephropathy subjects revealed several low-abundance proteins that reproducibly distinguished low glomerular filtration rate (GFR) from both high GFR diabetic and normal subjects, including uteroglobin, a protein previously associated with renal scarring. Conclusions and clinical relevance: We developed a novel method to identify low-abundance urinary proteins that enables the discovery of potential biomarkers to improve the diagnosis and management of patients with diabetic nephropathy.

Original languageEnglish (US)
Pages (from-to)603-612
Number of pages10
JournalProteomics - Clinical Applications
Issue number11-12
StatePublished - Dec 2011


  • Diabetic nephropathy
  • Immunodepletion
  • LC
  • Proteomic methods
  • Urinary biomarkers

ASJC Scopus subject areas

  • Clinical Biochemistry


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