Proteomic detection of prostate-specific antigen using a serum fractionation procedure: potential implication for new low-abundance cancer biomarkers detection.
One of the major obstacles in proteomic analysis of biological fluids is the presence of highly abundant proteins such as albumin and immunoglobulins, which can interfere with the resolution and sensitivity of the proteome profiling techniques used. In this paper, we describe an anion exchange fractionation procedure for serum using denaturating conditions allowing protein-protein interaction disruption before analysis by surface-enhanced laser desorption/ionization and by two-dimensional electrophoresis. This method simplifies the serum proteome into subproteomes and markedly increases resolution and sensitivity without any loss of minor proteins. To confirm the applicability of this method, fractionated serum of a patient with prostate cancer was analyzed for the presence of the prostate-specific antigen (PSA) which is a low-abundance tumor marker protein. The results demonstrate that PSA can be detected by two-dimensional electrophoresis only in serum following fractionation. Hence, this procedure may facilitate the identification of other, so far unknown, tumor markers in patient sera.
Solassol
Jérôme
J
Hôpital Arnaud de Villeneuve, INSERM U540, 191 Avenue du Doyen Giraud, 34295 Montpellier Cedex 5, France. j-solassol@chu-montpellier.fr
Marin
Philippe
P
Demettre
Edith
E
Rouanet
Philippe
P
Bockaert
Joël
J
Maudelonde
Thierry
T
Mangé
Alain
A
D000818:Animals; D015415:Biomarkers / Q000097:blood*; D005591:Chemical Fractionation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007474:Ion Exchange; D013058:Mass Spectrometry; D017430:Prostate-Specific Antigen / Q000097:blood*; D040901:Proteomics / Q000379:methods
D015415:Biomarkers; D017430:Prostate-Specific Antigen