Urinary Proteome Profiling Using 2D-DIGE and LC-MS/MS - LC-MS/MS in Proteomics: Methods and Applications - page 290

Chapter 18

Urinary Proteome Profiling Using 2D-DIGE and LC-MS/MS

Mark E. Weeks

Abstract

Proteomic methodologies have been at the forefront of cancer research for several years. The use of
proteomic strategies to study all expressed genes aims to discover biomarkers indicative of the physio-
logical state of cancer cells at specific time points, enabling early diagnosis, following cancer develop-
ment/progression, screening and monitoring the efficacy of new therapeutic agents. Onco-proteomics
has the potential to impact on oncology practice by delivering individualised highly selective clinical care.
2D-DIGE (2D difference in gel electrophoresis) enables simultaneous examination and comparison of
multiple samples using cyanine dyes to label amino acid residues that are then separated based on charge
and mass. These advantages combined with universal availability have until recently made 2D-DIGE a
first method of choice in cancer proteome analysis of diverse specimens, including tissues, cell lines, blood
and other body fluids.

Key words: Proteomics, 2D-DIGE, urine, cancer, HPLC-Chip/MS system, LC-MS/MS.

1. Introduction

Proteomics encompasses the identification of changes in protein
expression, sub-cellular distribution, post-translational modifica-
tions and the deciphering of protein-protein interactions. Onco-
proteomics aims to isolate, identify and recognise patterns of
protein expression that differentiate different physiological states
(cancer vs benign, early cancer vs advanced cancer states, etc.)
( 1 ) . Pancreatic ductal adenocarcinoma accounts for over 213,000
deaths worldwide each year, largely due to late presentation and
diagnosis ( 2 ) . Preliminary studies demonstrate that urine can be a
valid source for non-invasive biomarker discovery in patients with
pancreatic diseases ( 3 ) .