? Urine is an easily accessed specimen that has not been well exploited for cancer biomarker studies. We have developed a new stable isotope mass tag (13C Phenylisocyanate, PIC) that has several advantages for protein quantification in complex mixtures. We describe a complete analytic platform for urine biomarker discovery using PIC labeling. This platform includes: 1) custom designed software (PICquant) to automatically quantify labeled peaks, 2) a spectrum-comparison algorithm that groups spectra into a Registry of spectra representing unique peptide families, and 3) enhanced peptide sequencing by distinguishing b- and y-ion series in CID spectra. Completing the platform is a clinical registry that links acquired specimens to current and prospective clinical information including outcomes, and that enables multivariate clustering of disease states with quantified protein families. The mature PICquant platform will provide nearly completely automated data analysis, allowing assembly of numerous patient samples into complete protein abundance profiles akin to gene expression array data. Additionally, the expanding Registry database will be of use to other investigators processing either PIC-labeled or unlabeled peptide spectra. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA126101-02
Application #
7294337
Study Section
Special Emphasis Panel (ZCA1-SRRB-9 (O1))
Program Officer
Rodriguez, Henry
Project Start
2006-09-27
Project End
2010-08-31
Budget Start
2007-09-05
Budget End
2008-08-31
Support Year
2
Fiscal Year
2007
Total Cost
$294,213
Indirect Cost
Name
University of Virginia
Department
Pathology
Type
Schools of Medicine
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Mondal, Arindam; Victor, Ken G; Pudupakam, R S et al. (2014) Newly identified phosphorylation site in the vesicular stomatitis virus P protein is required for viral RNA synthesis. J Virol 88:1461-72
Victor, Ken G; Rady, Joshua M; Cross, Janet V et al. (2012) Proteomic profile of reversible protein oxidation using PROP, purification of reversibly oxidized proteins. PLoS One 7:e32527
Lyons, Charles E; Victor, Ken G; Moshnikov, Sergey A et al. (2011) PICquant: a quantitative platform to measure differential peptide abundance using dual-isotopic labeling with 12C6- and 13C6-phenyl isocyanate. Anal Chem 83:856-65
Victor, Ken G; Murgai, Meera; Lyons, Charles E et al. (2010) MAZIE: a mass and charge inference engine to enhance database searching of tandem mass spectra. J Am Soc Mass Spectrom 21:80-7