TY - JOUR
T1 - Development of a Targeted Urine Proteome Assay for kidney diseases
AU - Cantley, Lloyd G.
AU - Colangelo, Christopher M.
AU - Stone, Kathryn L.
AU - Chung, Lisa
AU - Belcher, Justin
AU - Abbott, Thomas
AU - Cantley, Jennifer L.
AU - Williams, Kenneth R.
AU - Parikh, Chirag R.
N1 - Publisher Copyright:
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2016/1/1
Y1 - 2016/1/1
N2 - 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.
AB - 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.
KW - Chronic kidney disease
KW - Polycystic kidney disease
KW - Targeted proteomics
KW - Urine
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U2 - 10.1002/prca.201500020
DO - 10.1002/prca.201500020
M3 - Article
C2 - 26220717
AN - SCOPUS:84953347455
SN - 1862-8346
VL - 10
SP - 58
EP - 74
JO - Proteomics - Clinical Applications
JF - Proteomics - Clinical Applications
IS - 1
ER -