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2015 ; 14
(6
): 1672-83
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An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based
Proteomic Profiling of Rare Cells in Whole Blood
#MMPMID25755294
Li S
; Plouffe BD
; Belov AM
; Ray S
; Wang X
; Murthy SK
; Karger BL
; Ivanov AR
Mol Cell Proteomics
2015[Jun]; 14
(6
): 1672-83
PMID25755294
show ga
Isolation and molecular characterization of rare cells (e.g. circulating tumor
and stem cells) within biological fluids and tissues has significant potential in
clinical diagnostics and personalized medicine. The present work describes an
integrated platform of sample procurement, preparation, and analysis for deep
proteomic profiling of rare cells in blood. Microfluidic magnetophoretic
isolation of target cells spiked into 1 ml of blood at the level of 1000-2000
cells/ml, followed by focused acoustics-assisted sample preparation has been
coupled with one-dimensional PLOT-LC-MS methodology. The resulting zeptomole
detection sensitivity enabled identification of ?4000 proteins with injection of
the equivalent of only 100-200 cells per analysis. The characterization of rare
cells in limited volumes of physiological fluids is shown by the isolation and
quantitative proteomic profiling of first MCF-7 cells spiked into whole blood as
a model system and then two CD133+ endothelial progenitor and hematopoietic cells
in whole blood from volunteers.