Current Gel Electrophoresis Approaches to Low-Abundance Protein Marker Discovery - Proteomic and Metabolomic Approaches to Biomarker Discovery - page 178

Native extracts CPLL-treated extracts

3 p 1 3 pH 1

5 p 8 5 pH 8

FIGURE 2 Real-life examples of reduction of dynamic concentration range upon CPLL treatment of two biological
samples: red blood cell lysate (upper panels) and human serum (lower panels). The left 2-DE images represent the spot
patterns of samples before the treatment; the right images evidence spot patterns after the peptide ligand library treatment.
The arrows represent the positioning of hemoglobin monomer before and after treatment; double arrows represent the
albumin spot in serum before and after the treatment.

electrophoresis. Among upregulated proteins
fromovarianhyperstimulation syndrome patients
were ITI-H4, apolipoprotein A-IV, complement
factor I, and pigment epithelium-derived factor.
These potential biomarkers remain to be con-

of the synergistic effect of technologies described
in this chapter.
To resolve the high level of protein composi-
tion complexity of human serum after CPLL
treatment and in view of discovering expression
differences, elution fractionation was carried
out prior to 2-DE analysis. 49 The obtained frac-
tions showed complementary composition,
thus simplifying largely the interpretation of
2-DE patterns (see Figure 3 ). The suitability of
such an approach for potential biomarker detec-
tion was assessed with over 100 patients. The
maintenance of protein abundance was demon-
strated for C-reactive protein (a marker of
in

firmed and validated.
Plasma was used as a source of possible
biomarkers for hepatitis B associated with
cirrhosis 55 in a pilot study of eight selected
patients. Plasma treated with ProteoMiner

and analyzed by 2-DE showed spots of misregu-
lated proteins. One of them, clearly downregu-
lated, was identi c variant of the

gelsolin actin-depolymerizing factor. Although
further investigations are necessary to con

ed as a speci

rm
and validate, this was an interesting example

ammation) upon CPLL treatment. Reproduc-
ibility of the entire process from blood collection