Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification
Journal of Extracellular Vesicles,
Год журнала:
2024,
Номер
13(10)
Опубликована: Окт. 1, 2024
Abstract
Extracellular
vesicles
(EVs)
are
lipid
nanoparticles
and
play
an
important
role
in
cell‐cell
communications,
making
them
potential
therapeutic
agents
allowing
to
engineer
for
targeted
drug
delivery.
The
expanding
applications
of
EVs
next
generation
medicine
is
still
limited
by
existing
tools
scaling
standardized
EV
production,
single
tracing
analytics,
thus
provide
only
a
snapshot
tissue‐specific
cargo
information.
Here,
we
present
the
Snorkel‐tag,
which
have
genetically
fused
surface
marker
protein
CD81,
series
tags
with
additional
transmembrane
domain
be
displayed
on
surface,
resembling
snorkel.
This
system
enables
affinity
purification
from
complex
matrices
non‐destructive
form
while
maintaining
characteristics
terms
profiles,
associated
miRNA
patterns
uptake
into
model
cell
line.
Therefore,
consider
Snorkel‐tag
widely
applicable
tool
research,
efficient
preparation
standards
reference
materials,
or
dissecting
different
markers
when
fusing
other
tetraspanins
vitro
vivo.
Язык: Английский
A cautionary note on the potential pitfalls of using N-terminal truncated CD63 to label small extracellular vesicles
Scientific Reports,
Год журнала:
2025,
Номер
15(1)
Опубликована: Март 1, 2025
Abstract
Small
extracellular
vesicles
(sEV)
are
nanosized
that
facilitate
intracellular
communication.
A
significant
research
obstacle
is
the
isolation
of
sEV
devoid
non-sEV
contaminants.
Immunoaffinity
capture
with
sEV-specific
antibodies
an
attractive
approach
to
purifying
sEV,
but
it
risks
disrupting
during
antibody
dissociation.
Furthermore,
immunoaffinity
may
require
modification
EV-specific
proteins
for
incorporation
tags
on
EV
surface,
unknown
implications
production
and
function.
The
aim
this
study
was
investigate
whether
a
previously
reported
CD63
truncation
efficient
small
extravesicular
surface.
We
therefore
conjugated
ALFA-tag
N-terminal-truncated
CD63,
included
nanoluciferase
at
C-terminus,
luminescent
tracing
sEV.
Full-length
CD63-nanoluciferase
used
as
control.
Plasmid
constructs
expressing
these
were
transfected
into
HEK293
cells.
In
contrast
previous
report,
N-terminal
impaired
its
membrane
localisation
reduced
yield
EVs.
Further
investigation
revealed
some
tagged
co-localized
aggresomes
preferentially
secreted
from
cells
soluble
protein
rather
than
being
associated
These
results
demonstrate
can
impair
function
yield,
potentially
generating
misleading
results.
Язык: Английский
Cutting‐Edge Aptasensing Approaches for Electrochemical Detection of Exosomes Utilizing MXenes
ChemElectroChem,
Год журнала:
2025,
Номер
unknown
Опубликована: Апрель 1, 2025
Herein,
the
basic
properties,
composition,
and
isolation
methods
for
separation
of
exosomes
are
described,
since
they
can
be
a
rich
source
disease
biomarkers.
Then
an
introduction
to
MXenes,
novel
class
2D
nanomaterials
with
interesting
properties
applicable
in
numerous
fields
including
biosensing
is
provided.
Also,
aptamers
described
as
alternative
antibodies
robust
biorecognition
analytes
interest.
The
final
part
article
gives
examples
which
these
three
key
components
integrated
sensitive
and/or
electrochemical
detection
exosomes.
conclusion
provides
summary
initial
achievements
also
outlook
future
discoveries
exosome
aptasensing
using
advanced
nanomaterials,
i.e.,
MXenes.
MXenes
have
promising
affinity‐based
biosensing,
being
hydrophilic
surface
functional
groups.
In
addition,
free
plasmons
present
used
covalent
grafting
elements
diazonium
moieties.
This
especially
approach
immobilization
DNA/RNA
aptamers,
readily
modified
by
Язык: Английский
Isolation and characterization of bone mesenchymal cell small extracellular vesicles using a novel mouse model
Journal of Bone and Mineral Research,
Год журнала:
2024,
Номер
39(11), С. 1633 - 1643
Опубликована: Авг. 22, 2024
Abstract
Extracellular
vesicles
(EVs)
are
key
mediators
of
cell–cell
communication
and
involved
in
transferring
specific
biomolecular
cargo
to
recipient
cells
regulate
their
physiological
functions.
A
major
challenge
the
understanding
EV
function
vivo
is
difficulty
ascertaining
origin
particles.
The
recent
development
“Snorkel-tag,”
which
includes
EV-membrane-targeted
CD81
fused
a
series
extra-vesicular
protein
tags,
can
be
used
mark
EVs
originating
from
source
for
subsequent
isolation
characterization.
We
developed
an
mouse
model,
termed
“CAGS-Snorkel,”
expresses
Snorkel-tag
under
control
Cre-lox
system,
crossed
this
with
either
Prx1-Cre
(mesenchymal
progenitors)
or
Ocn-Cre
(osteoblasts/osteocytes)
isolated
Snorkel-tagged
bone
marrow
plasma
using
magnetic
bead
affinity
column.
miRNA-sequencing
was
performed
on
EVs,
although
similar
profiles
were
observed,
few
miRNAs
metabolism
(miR-106b-5p,
miRs-19b-3p,
miRs-219a-5p)
enriched
Ocn-derived
relative
Prx1-derived
subpopulations.
To
characterize
effects
these
small
cell
target,
cultured
stromal
treated
Prx1
Ocn
mRNA-sequencing
performed.
Pathways
ossification,
development,
extracellular
matrix
interactions
regulated
by
both
subpopulations,
whereas
pathways
including
advanced
glycation
end-products
signaling
uniquely
subpopulation,
underlying
important
biological
subpopulations
within
microenvironment.
These
data
demonstrate
that
CAGS-Snorkel
model
useful
tool
characterizing
biology
tissue-specific
EVs.
Moreover,
while
mesenchymal
populations
share
common
secretory
profile,
we
uncover
differences
based
stage
osteoblastic
differentiation
may
have
consequences.
Язык: Английский
A comprehensive summary of the ASEV-CzeSEV joint meeting on extracellular vesicles
Extracellular Vesicles and Circulating Nucleic Acids,
Год журнала:
2024,
Номер
5(4), С. 765 - 84
Опубликована: Дек. 21, 2024
This
report
summarizes
the
ASEV-CzeSEV
Joint
Meeting
on
Extracellular
Vesicles
(EVs),
held
at
Medical
University
of
Vienna
in
September
2024.
The
conference
focused
introducing
and
expanding
EV
research
infrastructure
within
Czech
Republic
Austria,
highlighting
areas
for
collaboration.
Key
sessions
featured
EV-based
diagnostics,
tissue
regeneration,
interspecies
communication
therapeutic
applications,
with
an
emphasis
shared
resources
cross-border
partnerships.
program
included
oral
poster
presentations
engineering,
new
isolation
techniques,
potential
clinical
as
well
industry
updates
latest
technologies.
meeting
concluded
awards
outstanding
reflecting
quality
work
presented.
Following
conference,
a
dedicated
workshop
was
flow
cytometry
analysis
EVs,
allowing
participants
to
deepen
their
technical
expertise
characterization.
captures
main
discussions,
findings,
collaborative
opportunities
explored
meeting,
signaling
strong
regional
support
advancing
research.
Язык: Английский