Journal of Medicinal Chemistry,
Год журнала:
2024,
Номер
unknown
Опубликована: Дек. 16, 2024
Radiolabeled
antibodies
are
promising
for
targeted
cancer
imaging,
but
their
structural
integrity
may
suffer
during
bioconjugation
and
radiolabeling,
leading
to
undetected
aggregation.
This
study
evaluates
dynamic
light
scattering
(DLS)
as
a
complementary
method
size-exclusion
high-performance
liquid
chromatography
(SEC-HPLC)
detecting
aggregation
in
radiolabeled
antibodies.
Trastuzumab
was
conjugated
with
NOTA
bifunctional
chelator
at
various
ratios,
[64Cu]CuCl2,
analyzed
by
using
DLS
SEC-HPLC
before
after
purification.
revealed
significant
preparation,
SEC-HPLC,
showed
reduced
aggregates
following
Tumor-targeting
efficacy
correlated
intact
antibody
content,
Pearson's
correlations
of
0.71
(PET
imaging)
0.75
(biodistribution)
NIH3T6.7
tumor-bearing
mice.
The
findings
suggest
vital
quality
control
tool,
offering
enhanced
detection
By
adopting
DLS,
the
bioactivity
can
be
better
predicted,
potentially
improving
reliability
effectiveness
these
radiopharmaceuticals
clinical
settings.
Analytical Chemistry,
Год журнала:
2025,
Номер
unknown
Опубликована: Март 21, 2025
Monoclonal
antibodies
(mAbs)
are
widely
used
in
tumor
and
autoimmune
disease
therapy
clinical
diagnosis,
but
they
suffer
from
self-aggregation,
particularly
dimer
formation,
impacting
their
stability,
efficacy,
potentially
causing
severe
allergies.
Traditional
methods
for
detecting
mAb
dimers,
such
as
TEM,
AUC,
DLS,
SEC,
CE,
limited
by
low
throughput,
high
costs,
qualitative
data,
making
them
unsuitable
large-scale
sample
assays
biopharmaceuticals
industry
well
antibody
research.
To
address
these
issues,
we
developed
a
facile
method
of
protein
cross-linking
gel
electrophoresis
with
online
intrinsic
fluorescence
imaging
(PC-GE-IFI)
self-aggregation
stability
mAbs.
This
enables
the
real-time
quantification
monoclonal
(mAb)
monomers
dimers
exceptional
sensitivity,
characterized
detection
limits
(monomer:
0.9
nM;
dimer:
0.45
nM)
broad
dynamic
range
2.50-2500
1.25-1250
nM).
Furthermore,
wells
can
be
windows
assay
precipitates
formed
aggregation.
The
supports
accurate
assessment
dimerization
monomer
purity
under
various
stress
conditions,
including
thermal
stress,
mechanical
agitation,
freeze-thaw
cycles.
Moreover,
allows
concurrent
analysis
across
multiple
samples
at
different
concentrations,
while
nonlinear
fitting
provides
dissociation
constant
(Kd)
monomer-dimer
interactions,
critical
parameter
that
aids
assessing
aggregation
propensity,
informs
design
development
products
PC-GE-IFI
has
great
potential
development,
quality
control,
safety
mAbs,
bispecific
antibodies,
ADCs,
drugs.
Analytical Chemistry,
Год журнала:
2024,
Номер
96(28), С. 11155 - 11162
Опубликована: Июнь 29, 2024
Protein-based
biopharmaceutical
drugs,
such
as
monoclonal
antibodies,
account
for
the
majority
of
best-selling
drugs
globally
in
recent
years.
For
bioprocesses,
key
performance
indicators
are
concentration
and
aggregate
level
product
being
produced.
In
water
NMR
(wNMR),
use
transverse
relaxation
rate
[R2(1H2O)]
has
been
previously
used
to
determine
protein
level;
however,
it
cannot
be
separate
between
them
without
using
an
additional
technique.
This
work
shows
that
is
possible
"decouple"
these
two
characteristics
by
recording
diffusion
coefficient
[D(1H2O)]
conjunction
with
R2(1H2O),
even
event
overlap
either
D(1H2O)
or
R2(1H2O).
method
demonstrated
on
three
different
systems,
following
appropriate
R2(1H2O)
calibration
data
acquisition
a
interest.
Our
highlights
potential
benchtop
at-line
process
analytical
