Dual-Functional Nanobody Optical Probes for In Vivo Fluorescence Imaging and Photodynamic Therapy
ACS Applied Materials & Interfaces,
Год журнала:
2025,
Номер
unknown
Опубликована: Фев. 14, 2025
Nanobodies
have
gained
significant
attention
as
promising
tools
for
cancer
diagnostics
and
treatment
due
to
their
unique
ability
precisely
target
specific
cells.
However,
a
major
challenge
lies
in
the
site-specific
incorporation
of
multifunctional
molecules
into
nanobodies,
it
is
essential
link
these
manner
that
preserves
nanobody's
function
stability
while
retaining
desired
therapeutic
or
diagnostic
properties.
This
study
outlines
development
dual-functional
nanobody
optical
probes
enhanced
interventions.
We
designed
clickable
linker
enables
functionalization
nanobody,
facilitating
simultaneous
conjugation
two
dyes:
indocyanine
green
imaging
chlorin
e6
photodynamic
therapy.
In
vitro
cellular
assays
confirmed
successful
labeling
dyes,
with
probe
exhibiting
high
binding
specificity.
vivo
mice
bearing
Hep3B
tumors
revealed
clear
visualization
signal-to-noise
ratio.
Furthermore,
PEGylated
significantly
improved
tumor
retention,
enhancing
both
contrast
therapy
efficacy
compared
free
e6.
These
show
great
promise
precise
diagnosis
malignant
tumors.
Язык: Английский
Indium-111-Labeled Single-Domain Antibody for In Vivo CXCR4 Imaging Using Single-Photon Emission Computed Tomography
M. SPAHN,
Stephanie Mareike Anbuhl,
Kaat Luyten
и другие.
Bioconjugate Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Март 11, 2025
C-X-C
chemokine
receptor
type
4
(CXCR4)
is
highly
expressed
in
a
range
of
pathologies,
including
cancers
like
multiple
myeloma
and
non-Hodgkin
lymphoma,
inflammatory
diseases
such
as
rheumatoid
arthritis,
viral
infections
HIV.
Currently,
the
most
advanced
radiotracer
for
CXCR4
imaging
clinics
[68Ga]PentixaFor.
However,
its
structure
prone
to
modifications,
complicating
development
specific
fluorine-18-labeled
tracer
with
good
pharmacokinetic
properties.
This
study
aimed
screen
CXCR4-targeting
variable
domains
heavy-chain-only
antibody
(VHH
or
single-domain
(sdAb))
constructs
identify
promising
sdAb
vector
molecule
future
fluorine-18
tracer.
We
have
generated
five
CXCR4-specific
cysteine-containing
C-terminal
tag
(C-Direct
tag)
(VUN400-C-Direct,
VUN401-C-Direct,
VUN410-C-Direct,
VUN411-C-Direct,
VUN415-C-Direct)
one
probe
(VUN400-C)
without.
The
reduced
sdAbs
were
coupled
maleimide-DOTAGA
111In-labeling.
Their
binding
affinity
against
human
(hCXCR4)
was
assessed
by
using
previously
described
BRET-based
displacement
assay.
vivo
profile
naive
mice.
Based
on
plasma
stability
(60
min
post
injection
(p.i.)),
we
selected
VUN400-C-Direct
derivative
VUN400-C
further
evaluation.
These
compounds
([111In]In-DOTAGA-VUN400-C-Direct
[111In]In-DOTAGA-VUN400-C)
tested
mice
bearing
xenografts
derived
from
U87.CD4,
U87.CXCR4,
U87.CD4.CXCR4
cells
through
ex
biodistribution
studies
SPECT/CT
imaging.
six
labeled
high
radiochemical
conversion
(75–97%)
purity
(>95%).
In
radioactive
assays
cells,
[111In]In-DOTAGA-VUN400-C-Direct
[111In]In-DOTAGA-VUN401-C-Direct
displayed
highest
cellular
uptake,
achieving
10.4
±
1.6%
11.5
1.1%,
respectively.
mice,
showed
favorable
profile,
low
uptake
across
all
organs
except
kidneys
(Standardized
Uptake
Value
(SUV)
>
50,
n
=
3,
60
p.i.),
but
average
(40.6
9.4%,
p.i.).
xenografted
tumor
model,
only
minor
(SUVU87.CXCR4
0.71
0.002,
[111In]In-DOTAGA-VUN400-C
demonstrated
nearly
identical
(41.08
5.45%,
4)
CXCR4-expressing
(SUVU87.CD4.CXCR4
3.75
1.08
vs
SUVU87.CD4
0.64
0.19,
5,
which
could
be
blocked
coinjection
AMD3100
(5
mg/kg)
0.55
0.32
0.39
0.07,
2,
conclusion,
exhibited
vitro
hCXCR4.
Among
these,
properties,
indicating
VUN400-C's
potential
PET
applications
fluorine-18.
Язык: Английский
Development of [18F]AlF-RESCA-H006 as a Novel Molecular Probe for 5T4 Tumor Imaging and Exploration of Its Application in Pancreatic Adenocarcinoma
Molecular Pharmaceutics,
Год журнала:
2025,
Номер
unknown
Опубликована: Май 30, 2025
Pancreatic
adenocarcinoma
(PAAD)
is
an
extremely
lethal
cancer,
making
early
detection
and
precise
staging
essential
for
extending
survival.
Upregulation
of
5T4
mRNA
was
detected
in
bioinformatics
analysis
using
data
from
the
Cancer
Genome
Atlas
Genotype-Tissue
Expression.
In
current
work,
we
confirmed
overexpression
protein
tumor
samples
PAAD
compared
with
corresponding
paraneoplastic
tissue.
To
visualize
via
positron
emission
tomography
(PET)
noninvasively,
successfully
developed
[18F]AlF-RESCA-H006
based
on
a
single-domain
antibody
fragment.
The
radiotracer
demonstrated
high
binding
affinity
toward
antigens
vitro
remained
stable
final
formulation
up
to
4
h
storage
at
room
temperature.
two
preclinical
mouse
models
pancreatic
clearly
identified
sites,
showing
radioactivity
accumulation
2.43
±
0.46%
ID/g
Balb/C
mice
BxPC-3
xenografts
2.88
1.02%
NCG
PANC-1
xenografts.
Taken
together,
promising
PET
tracer
detecting
antigen
alteration
PAAD.
Further
investigations
are
required
assess
its
defluorination
level
higher
species
permeability
through
dense
extracellular
matrix
cancer.
Язык: Английский
Selection of the optimal chelator for labeling of DARPin Ec1 with gallium-68 for PET imaging of EpCAM expression
EJNMMI Radiopharmacy and Chemistry,
Год журнала:
2025,
Номер
10(1)
Опубликована: Май 30, 2025
Abstract
Background
Epithelial
cell
adhesion
molecule
(EpCAM)
is
a
transmembrane
glycoprotein,
which
overexpressed
in
several
types
of
malignancies.
Designed
ankyrin
repeat
protein
(DARPin)
Ec1
19
kDa
engineered
scaffold
that
binds
with
high
affinity
to
EpCAM.
Radiolabelled
might
be
used
as
companion
diagnostic
for
the
selection
patients
personalized
therapy.
This
study
aimed
investigate
influence
different
radiometal-chelator
complexes
on
biodistribution
and
imaging
contrast
68
Ga-labelled
Ec1.
To
this,
two
macrocyclic
chelators,
1,4,7-triazacyclononane-N,N,N-triacetic
acid
(NOTA)
1-(1,3-carboxypropyl)-1,4,7-triazacyclononane-4,7-diacetic
(NODAGA)
were
conjugated
C-terminus
The
previously
developed
DARPin
1,4,7,10-tetraazacylododecane-1,4,7,10-tetraacetic
(DOTA)
was
comparator.
Results
All
variants
successfully
labelled
Ga.
use
NOTA
NODAGA
provided
twice
higher
radiochemical
yield
improved
label
stability
compared
DOTA.
bound
EpCAM-expressing
cells
nanomolar
preserved
targeting
specificity
vitro
vivo.
Biodistribution
studies
mice
bearing
SKOV-3
xenografts
showed
[
Ga]Ga-Ec1-NOTA
had
lower
uptake
most
normal
organs
while
maintaining
tumor
uptake.
Among
all
variants,
lowest
liver
uptake,
no
significant
differences
Additionally,
highest
tumor-to-blood
ratio
Ga]Ga-Ec1-DOTA
Ga]Ga-Ec1-NODAGA.
Conclusion
preferred
radioconjugate
PET
EpCAM
expression.
Язык: Английский