Editorial Type:
Urology

Evaluation of mass spectrometry of urinary proteins and peptides as biomarkers for cats at risk of developing azotemia

Rosanne E. Jepson Department of Veterinary Clinical Sciences, Royal Veterinary College, NW1 0TU, London, England.

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 BVSc, MVetMed, PhD
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Gary R. Coulton St George's Medical Biomics Centre, St George's University of London, SW17 0RE, London, England.

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 PhD
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Matthew L. Cowan St George's Medical Biomics Centre, St George's University of London, SW17 0RE, London, England.

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Peter Markwell WALTHAM, a Division of Mars Inc, LE14 4RT, Melton Mowbray, England.

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Harriet M. Syme Department of Veterinary Clinical Sciences, Royal Veterinary College, NW1 0TU, London, England.

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 BVetMed, PhD
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Jonathan Elliott Department of Veterinary Basic Sciences, Royal Veterinary College, NW1 0TU, London, England.

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 VetMB, PhD
DOI:
https://doi.org/10.2460/ajvr.74.2.333
Volume/Issue:
Volume 74: Issue 2
Online Publication Date:
01 Feb 2013
Restricted access
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Abstract

Objective—To evaluate proteomic delineation of feline urine by mass spectrometry as a method for identifying biomarkers in cats at risk of developing azotemia.

Samples—Urine samples from geriatric cats (> 9 years old) with chronic kidney disease and nonazotemic cats that either remained nonazotemic (n = 10) or developed azotemia (10) within 1 year.

Procedures—Optimization studies with pooled urine were performed to facilitate the use of surface enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) for analysis of the urinary proteome of cats. Urine samples from nonazotemic cats at entry to the study were analyzed via SELDI-TOF-MS with weak cation exchange and strong anion exchange arrays. Spectral data were compared to identify biomarkers for development of azotemia.

Results—Low protein concentration in feline urine precluded direct application to array surfaces, and a buffer exchange and concentration step was required prior to SELDI-TOF-MS analysis. Three preparation conditions by use of weak cation and strong anion exchange arrays were selected on the basis of optimization studies for detection of biomarkers. Eight potential biomarkers with an m/z of 2,822, 9,886, 10,033, 10,151, 10,234, 11,653, 4,421, and 9,505 were delineated.

Conclusions and Clinical Relevance—SELDI-TOF-MS can be used to detect urinary low-molecular weight peptides and proteins that may represent biomarkers for early detection of renal damage. Further study is required to purify and identify potential biomarkers before their use in a clinical setting.

Abstract

Objective—To evaluate proteomic delineation of feline urine by mass spectrometry as a method for identifying biomarkers in cats at risk of developing azotemia.

Samples—Urine samples from geriatric cats (> 9 years old) with chronic kidney disease and nonazotemic cats that either remained nonazotemic (n = 10) or developed azotemia (10) within 1 year.

Procedures—Optimization studies with pooled urine were performed to facilitate the use of surface enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) for analysis of the urinary proteome of cats. Urine samples from nonazotemic cats at entry to the study were analyzed via SELDI-TOF-MS with weak cation exchange and strong anion exchange arrays. Spectral data were compared to identify biomarkers for development of azotemia.

Results—Low protein concentration in feline urine precluded direct application to array surfaces, and a buffer exchange and concentration step was required prior to SELDI-TOF-MS analysis. Three preparation conditions by use of weak cation and strong anion exchange arrays were selected on the basis of optimization studies for detection of biomarkers. Eight potential biomarkers with an m/z of 2,822, 9,886, 10,033, 10,151, 10,234, 11,653, 4,421, and 9,505 were delineated.

Conclusions and Clinical Relevance—SELDI-TOF-MS can be used to detect urinary low-molecular weight peptides and proteins that may represent biomarkers for early detection of renal damage. Further study is required to purify and identify potential biomarkers before their use in a clinical setting.

Contributor Notes

The study was performed at the Royal Veterinary College and the St George's Medical Biomics Centre, St. George's University of London.

Supported by Mars Veterinary.

Address correspondence to Dr. Jepson (rjepson@rvc.ac.uk).
Cover American Journal of Veterinary Research
American Journal of Veterinary Research
Publication Date:
01 Feb 2013

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