TY - JOUR
T1 - 2D-electrophoresis and the urine proteome map
T2 - Where do we stand?
AU - Candiano, Giovanni
AU - Santucci, Laura
AU - Petretto, Andrea
AU - Bruschi, Maurizio
AU - Dimuccio, Veronica
AU - Urbani, Andrea
AU - Bagnasco, Serena
AU - Ghiggeri, Gian Marco
PY - 2010/3/10
Y1 - 2010/3/10
N2 - The discovery of urinary biomarkers is a main topic in clinical medicine. The development of proteomics has rapidly changed the knowledge on urine protein composition and probably will modify it again. Two-dimensional electrophoresis (2D-PAGE) coupled with mass spectrometry has represented for years the technique of choice for the analysis of urine proteins and it is time to draw some conclusions. This review will focus on major methodological aspects related to urine sample collection, storage and analysis by 2D-PAGE and attempt to define an advanced normal urine protein map. Overall, 1118 spots were reproducibly found in normal urine samples but only 275 were characterized as isoforms of 82 proteins. One-hundred height spots belonging to 30 proteins were also detected in plasma and corresponded to typical plasma components. The identity of most of the proteins found in normal urine by 2D-PAGE remains to be determined, the majority being low-molecular weight proteins (< 30 kDa). Equalization procedures would also enhance sensitivity of the analysis and allow low abundance proteins to be characterized. Therefore, we are still on the way to define the normal urine composition. Technology advancements in concentrating procedure will improve sensitivity and give the possibility to purify proteins for mass spectrometry.
AB - The discovery of urinary biomarkers is a main topic in clinical medicine. The development of proteomics has rapidly changed the knowledge on urine protein composition and probably will modify it again. Two-dimensional electrophoresis (2D-PAGE) coupled with mass spectrometry has represented for years the technique of choice for the analysis of urine proteins and it is time to draw some conclusions. This review will focus on major methodological aspects related to urine sample collection, storage and analysis by 2D-PAGE and attempt to define an advanced normal urine protein map. Overall, 1118 spots were reproducibly found in normal urine samples but only 275 were characterized as isoforms of 82 proteins. One-hundred height spots belonging to 30 proteins were also detected in plasma and corresponded to typical plasma components. The identity of most of the proteins found in normal urine by 2D-PAGE remains to be determined, the majority being low-molecular weight proteins (< 30 kDa). Equalization procedures would also enhance sensitivity of the analysis and allow low abundance proteins to be characterized. Therefore, we are still on the way to define the normal urine composition. Technology advancements in concentrating procedure will improve sensitivity and give the possibility to purify proteins for mass spectrometry.
KW - DIGE
KW - Mass spectrometry
KW - Normal urine
KW - Solid-phase ligand library
KW - Two-dimensional electrophoresis
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U2 - 10.1016/j.jprot.2009.12.003
DO - 10.1016/j.jprot.2009.12.003
M3 - Review article
C2 - 20004755
AN - SCOPUS:77249137641
SN - 1874-3919
VL - 73
SP - 829
EP - 844
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 5
ER -