72 / 120
Liposomes, Exosomes, and Virosomes: From Modeling Complex
Membrane Processes to Medical Diagnostics and Drug Delivery
Poster Abstracts
67
46-POS
Board 23
On Chip Biodetection, Analysis and Nanometrology of Cell-Derived Vesicles
Sameh OBEID
1
, Gaël Belliot
2
, Benoit Le Roy de Boiseaumarié
1
, Adam Ceroi
3
, Guillaume
Mourey
3
, Philippe Saas
3
, Wilfrid Boireau
1
, Céline Elie-Caille
1
.
1
FEMTO-ST Institute, Besançon, France,
2
National Reference Center for Enteric Viruses, Public
Hospital, Dijon, France,
3
INSERM U1098, EFS Bourgogne Franche-Comté, UBFC, Besançon,
France.
Extracellular vesicles (EVs) are small vesicles (10-4000 nm) shed from different cells types upon
activation or apoptosis. EVs, including exosomes, microparticles and apoptotic bodies, are
recognized to play an important role in various biological processes and are also recognized as
potential biomarkers of various health disorders
1
.
Currently, the technic of choice used for the detection of EVs is flow cytometry (FC)
2
, but the
main drawback of FC is the lower detection limit (≈300nm at best); consequently, a big fraction
of vesicles cannot be detected.
Our work aims to use the potential of surface plasmon resonance (SPR) method to detect, and
qualify, specifically and without labeling, the EVs in their complex media, via their
(immune)capture onto the biochip-surface using different specific ligands.
Using two calibration standards of known size and concentration (Virus-Like Particles “VLP”
(50nm) & modified melamin resin MF beads (920nm)), we have shown that the capture level of
these entities by their specific ligands is linearly related to their concentration in the sample.
Dynamic ranges of concentrations (8x10
10
to 2x10
12
VLPs/ml and 3,25x10
7
to 3,25x10
8
beads/ml) have been established for 2 min injection time. These concentration ranges match the
typical range of EVs natural concentrations in plasma
3,4
. Additionally, by varying the injection
time, the dynamic range increases, which provides more sensibility for the detection of MPs in a
wide concentration range.
Moreover, a combination between SPR and atomic force microscopy (AFM) approaches appears
to be suitable for the metrological analysis of captured EVs, which allows identification of their
size subpopulations. Furthermore, an on-chip proteomic study was also engaged to discover
specific proteomic profiling of EVs in different conditions.
At last, we have adjusted this analytical platform based on a gold biochip to qualify and compare
resting and activated platelet MPs.