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Proteomics Clin Appl. 2016 Jan;10(1):58-74. doi: 10.1002/prca.201500020. Epub 2015 Oct 6.

Development of a Targeted Urine Proteome Assay for kidney diseases.

Author information

1
Department of Internal Medicine, School of Medicine, Yale University, New Haven, CT, USA.
2
Primary Ion, Old Lyme, CT, USA.
3
W.M. Keck Foundation Biotechnology Laboratory, School of Medicine, Yale University, New Haven, CT, USA.
4
Department of Molecular Biophysics and Biochemistry, School of Medicine, Yale University, New Haven, CT, USA.
5
Program of Applied Translational Research, Yale University School of Medicine, New Haven, CT, USA.

Abstract

PURPOSE:

Since human urine is the most readily available biofluid whose proteome changes in response to disease, it is a logical sample for identifying protein biomarkers for kidney diseases.

EXPERIMENTAL DESIGN:

Potential biomarkers were identified by using a multiproteomics workflow to compare urine proteomes of kidney transplant patients with immediate and delayed graft function. Differentially expressed proteins were identified, and corresponding stable isotope labeled internal peptide standards were synthesized for scheduled MRM.

RESULTS:

The Targeted Urine Proteome Assay (TUPA) was then developed by identifying those peptides for which there were at least two transitions for which interference in a urine matrix across 156 MRM runs was <30%. This resulted in an assay that monitors 224 peptides from 167 quantifiable proteins.

CONCLUSIONS AND CLINICAL RELEVANCE:

TUPA opens the way for using a robust mass spectrometric technology, MRM, for quantifying and validating biomarkers from among 167 urinary proteins. This approach, while developed using differentially expressed urinary proteins from patients with delayed versus immediate graft function after kidney transplant, can be expanded to include differentially expressed urinary proteins in multiple kidney diseases. Thus, TUPA could provide a single assay to help diagnose, prognose, and manage many kidney diseases.

KEYWORDS:

Chronic kidney disease; Polycystic kidney disease; Targeted proteomics; Urine

PMID:
26220717
PMCID:
PMC5003777
DOI:
10.1002/prca.201500020
[Indexed for MEDLINE]
Free PMC Article

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