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Abstract

Biochemistry

Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis

Published: May 8th, 2021

DOI:

10.3791/62512

1Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Graecia University of Catanzaro, 2Department of Health Science, Magna Graecia University of Catanzaro, 3Romolo Hospital, 4Department of Experimental and Clinical Medicine, Magna Graecia University of Catanzaro

Abstract

Filter-aided sample protocol (FASP) is widely used for proteomics sample preparation because it allows to concentrate diluted samples and it is compatible with a wide variety of detergents. Bottom-up proteomics workflows like FASP increasingly rely on LC-MS/MS methods performed in data-independent analysis (DIA) mode, a scanning method that allows deep proteome coverage and low incidence of missing values.

In this report, we will provide the details of a workflow that combines a FASP protocol, a double StageTip purification step and LC-MS/MS in DIA mode for urinary proteome mapping. As a model sample, we analyzed expressed prostatic secretions (EPS)-urine, a sample collected after a digital rectal exam (DRE), which is of interest in prostate cancer biomarker discovery studies.

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Keywords Proteomic Profile

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