Biology Reference
In-Depth Information
Syringe Pump
Microinjection Valve
CITP Capillary
+
1
2
6
G
5
3
4
Waste
Trap Column
Trap Column
Trap Column
Trap Column
Microselection
Valve
1
1
6
2
2
6
5
3
3
5
4
4
Trap Column
Trap Column
Nano-
LC Pump
Database
Nano-RPLC Column
ESI-MS/MS
FIGURE 2
Schematic of online integration of CITP with nano-RPLC for achieving selective analyte enrichment and
multidimensional proteome separation. Solid and dashed lines represent the
flow paths for the loading of CITP fractions and
the injection of fractions into a nano-RPLC column, respectively.
Jinawath and colleagues
60
have applied the
CITP proteomic technology to perform compar-
ative proteomic analysis of paired primary and
recurrent post-chemotherapy ovarian high-
grade serous carcinomas from nine ovarian
cancer patients. The increase in ovarian cancer
proteome coverage, attributed to CITP-based
selective analyte enrichment, allowed the appli-
cation of protein network and pathway analysis
toward the discovery of ovarian carcinoma
biomarkers. For example, low-abundance pro-
teins such as cytokine IL-6 and signal transducer
and activator of transcription 3 (STAT3), as well
as many other proteins known to participate in
the IL-6 signaling pathway, have been identi
ed
and compared for their expression levels within
primary
and
recurrent
ovarian
tumors
(
Figure 4
).
The comparative proteomic
results have
further identi
ed RELA, which is the p65 subunit
of the NF-
B complex (
Figure 4
). p65 was overex-
pressed more than threefold in recurrent tumors
as compared to the primary tumors. The NF-
k
B/
RELA family of transcription factors is one of the
most important and well-characterized signaling
pathways in both normal and pathological
conditions. It controls a variety of cellular func-
tions
k
including in
ammatory and immune
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