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Enabling Sensitive Phenotypic Profiling of Cancer-Derived Small Extracellular Vesicles Using Surface-Enhanced Raman Spectroscopy Nanotags
journal contribution
posted on 2020-03-17, 13:47 authored by Wei Zhang, Lianmei Jiang, Russell J. Diefenbach, Douglas H. Campbell, Bradley J. Walsh, Nicolle H. Packer, Yuling WangCirculating
cancer-derived small extracellular vesicles (EVs) are
nanoscale membranous vesicles shed from cancer cells that are released
into surrounding body fluids. Small EVs contain biomolecules associated
with cancer such as DNA and proteins for cell-to-cell communication.
Therefore, small EVs have been regarded as important cancer biomarkers
for liquid biopsy-based cancer diagnosis and drug treatment monitoring.
However, because of the high heterogeneity and low level of small
EVs in body fluids, there is a high demand for sensitive detection
and characterization of such vesicles at a molecular level. In this
study, we have developed a sensitive and effective approach to simultaneously
profile multiple protein biomarkers expressed on cancer-derived small
EVs using surface-enhanced Raman spectroscopy (SERS) nanotags in a
single test, without complex isolation steps. Rapid and multiplexed
phenotypic profiling of small EVs is achieved by mixing specific detection
antibody-coated SERS nanotags, filtered conditioned EV-suspended medium
(conditioned EVs), and capture antibody (CD63)-conjugated magnetic
beads to form a sandwich immunoassay. As a proof-of-concept demonstration,
we applied this approach to characterize pancreatic cancer-derived
EVs by simultaneously detecting three specific EV surface receptors
including Glypican-1, epithelial cell adhesion molecules (EpCAMs),
and CD44 variant isoform 6 (CD44V6). The sensitivity of this method
was measured down to 2.3 × 106 particles/mL, which
is more sensitive and shows higher multiplexing capability than most
other reported EV profiling techniques, such as western blot, enzyme-linked
immunosorbent assay, and flow cytometry. Furthermore, phenotypic profiling
of small EVs from colorectal cancer and bladder cancer cell lines
(SW480 and C3) was conducted and compared to those derived from pancreatic
cancer (Panc-1), highlighting the significant difference in EV phenotypes
for various cancer cell types suspended in both phosphate-buffered
saline and plasma. Thus, we believe that this technology enables a
comprehensive evaluation of small secreted EV heterogeneity with high
sensitivity, offering strong potential for accurate noninvasive cancer
diagnosis and monitoring of drug treatment. In addition, this assay
provides point-of-care use because of the easy sample preparation
and portable nature of the Raman spectrometer.
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detection antibody-coated SERS nanotagspancreatic cancer-derived EVsbody fluids44VSurface-Enhanced Raman Spectroscopy Nanotags Circulating cancer-derivedenzyme-linked immunosorbent assaysurface-enhanced Raman spectroscopyEV surface receptorsDNAbladder cancer cell linesSWbiopsy-based cancer diagnosisnoninvasive cancer diagnosisnanoscale membranous vesiclesdrug treatment monitoringcancer cell typesCD 44 variant isoform 6
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