Journal of Separation Science,
Год журнала:
2024,
Номер
47(18)
Опубликована: Сен. 1, 2024
ABSTRACT
Multiple‐step
on‐line
preconcentration,
a
combination
of
at
least
two
stacking
techniques
has
been
developed
to
increase
the
sensitivity
in
capillary
electrophoresis
(CE)
for
analytes
various
samples.
It
is
usually
conducted
sequentially,
or
some
cases,
synergistically,
where
different
modes
occur
simultaneously.
allow
simultaneous
preconcentration
and
separation
kinds
complex
samples
single
CE
run.
This
review
aims
provide
recent
advances
multiple‐step
CE.
We
critically
technical
papers
published
last
7
years
up
until
July
2024,
subsequently
organized
according
main
techniques,
that
is,
field
amplification,
large
volume
sample
stacking,
transient
isotachophoresis,
micelle
solvent
cyclodextrin
others.
The
procedures,
fundamental
mechanism,
analytical
figures
merits
achieved,
their
feasibility
complicated
matrices
are
reviewed.
ABSTRACT
The
ICH
Q14
guideline
introduces
a
structured
framework
for
analytical
method
development
based
on
Analytical
Quality
by
Design
(AQbD)
principles,
aiming
to
ensure
robust,
reliable,
and
fit‐for‐purpose
methods
throughout
the
product
lifecycle.
However,
implementing
remains
challenging
due
lack
of
complete
examples
training
resources,
making
it
difficult
organizations
translate
theory
into
practice.
Although
previous
studies
have
applied
AQbD
capillary
electrophoresis
development,
many
focused
only
specific
aspects
such
as
design
experiments
(DoEs)
or
target
profile
(ATP),
leaving
gap
in
providing
comprehensive,
practical
tools
entire
This
manuscript
presents
novel,
user‐friendly
approach
AQbD,
offering
ready‐to‐implement
methodologies
that
simplify
process
design,
optimization,
validation,
implementation.
Through
stepwise
process,
provides
solutions
integrating
principles
everyday
workflows,
bridging
between
theoretical
concepts
real‐world
applications.
has
been
thoroughly
tested
diverse
industrial
settings,
demonstrating
its
reliability
effectiveness.
work
aims
facilitate
adoption
tools,
lessons
learned,
best
practices
align
with
guidelines.
Therapeutic
mAbs
show
a
specific
"charge
fingerprint"
that
may
affect
safety
and
efficacy,
and,
as
such,
it
is
often
identified
critical
quality
attribute
(CQA).
Capillary
iso-electric
focusing
(cIEF),
commonly
used
for
the
evaluation
of
such
CQA,
provides
an
analytical
tool
to
investigate
mAb
purity
identity
across
product
lifecycle.
Here,
we
discuss
results
analysis
panel
antibody
products
by
conventional
whole-column
imaging
cIEF
systems
performed
part
European
Pharmacopoeia
activities
related
development
"horizontal
standards"
control
monoclonal
antibodies
(mAbs).
The
study
aimed
at
designing
verifying
independent
transversal
procedure
reliable
charge
variants.
Despite
use
comparable
experimental
conditions,
discrepancies
in
profile
measured
isoelectric
points
emerged
between
two
systems.
These
data
suggest
are
method-dependent
rather
than
absolute,
aspect
known
experts
field
pharmaceutical
industry,
but
not
suitably
documented
literature.
Critical
implications
from
regulatory
perspectives,
herein
thoughtfully
discussed,
with
special
focus
on
context
market
surveillance
identification
falsified
medicines.
Journal of Pharmaceutical Analysis,
Год журнала:
2024,
Номер
14(12), С. 101004 - 101004
Опубликована: Май 23, 2024
Extracellular
vesicles
(EVs)
are
a
part
of
cell-to-cell
communication
system
prokaryotic
and
eukaryotic
organisms.
Their
ability
to
penetrate
biological
barriers
transfer
molecules
between
cells
shows
their
potential
as
novel
class
drug
delivery
platform.
However,
because
the
great
heterogeneity
EVs
complexity
matrices
from
which
they
typically
isolated,
reliable
quality
control
procedures
need
be
established
ensure
safety
for
medical
use.
According
current
recommendations,
quantification
EVs,
confirmation
identity,
purity
assessment
require
use
several
analytical
techniques,
including
particle-size
distribution
analysis,
proteomics,
electron
microscopy,
making
characterization
process
demanding.
Capillary
electrophoresis
(CE)
has
recently
emerged
an
alternative
tool
EV
characterization.
In
this
study,
available
literature
on
concept
was
reviewed.
Its
performance
critically
evaluated
compared
with
currently
used
methods.
The
utility
CE
in
EV-based
medicines
discussed.
ABSTRACT
Hemoglobinopathies,
hereditary
disorders
affecting
the
structure
or
production
of
hemoglobin,
were
detected
by
routine
HbA
1c
measurements
capillary
electrophoresis
(CE)
at
University
Hospital
Motol,
Prague.
The
potential
ultraviolet–visible
(UV–Vis)
and
Fourier‐transform
infrared
(FTIR)
spectroscopy
for
detection
characterization
hemoglobinopathies
was
investigated.
FTIR
spectra
recorded
with
a
very
high
resolution
(0.5
cm
−1
)
128
scans.
broad
amide
I
peak,
located
1700–1600
,
can
be
formed
superimposition
conformational
structures
hemoglobin.
These
secondary
protein
subjected
to
mathematical
analysis.
application
band
narrowing
techniques,
followed
curve
fitting
integration
processes,
provided
basis
quantitative
estimation
structure.
As
result,
unambiguous
differences
in
UV–Vis
among
patients
presumably
normal
an
HbC
hemoglobin
S/hemoglobin
G
(HbS/HbG)‐Philadelphia
variant
could
not
demonstrated.
However,
indicated
slight
α‐helix,
β‐turns,
β‐sheet,
random
coil
these
mutations.
In
spectral
wavenumber
range
950–850
there
some
obvious
specific
wavenumbers
between
those
variant.
Further
investigations
are
needed
sufficient
number
variants
elucidate
potency
hemoglobinopathies.
ABSTRACT
Biological
drugs
like
monoclonal
antibodies
require
careful
analysis
and
characterization
to
ensure
product
quality,
safety,
efficacy.
Charge
variants
of
the
molecule
are
key
interest
analyzed
using
imaged
capillary
isoelectric
focusing
(icIEF).
However,
deeper
these
poses
challenges.
Two
workflows
for
their
exist:
an
ion‐exchange
chromatography
method
variant
collection
before
mass
spectrometry
(MS)
analysis,
which
is
labor‐intensive,
direct
coupling
CE
MS,
allows
detailed
structural
but
has
limitations,
example,
due
incompatibilities
with
ES
ionization
high
BGE
concentrations.
This
study
evaluates
a
platform
that
fractionates
charge
offline
MS
analysis.
The
suitability
procedure
in
analytical
icIEF
methods
converted
into
preparative
cIEF
fractionation
by
increasing
sample
concentration
adding
20
mM
arginine
as
cathodic
spacer
was
tested.
After
chemical
mobilization
fraction
collection,
identity
fractions
determined
fluorescence
measurement
reinjection
protein‐containing
fractions,
previously
developed
method.
subsequently
performed.
general
workflow
demonstrated
Matuzumab.
Transferring
from
0.2
1.2
mg/mL
yielded
identical
number
peaks
visually
comparable
peak
profiles.
separation
took
50
min,
additional
25
min
45
s
per
totaling
approximately
2.5
h.
Verification
isolation
straightforward
via
icIEF.
Following
fractionation,
allowed
identification
main
peaks.
Preliminary
results
other
indicated
range
experiments
needs
further
investigation.
Future
work
will
aim
optimize
sensitivity,
selectivity,
time,
reproducibility.
Frontiers in Chemistry,
Год журнала:
2025,
Номер
13
Опубликована: Фев. 20, 2025
Biopharmaceuticals
are
increasingly
utilised
in
the
treatment
of
oncological,
inflammatory,
and
autoimmune
diseases,
largely
due
to
their
exceptional
specificity
targeting
antigens.
However,
structural
complexity,
heterogeneity,
sensitivity
pose
crucial
challenges
production,
purification,
delivery.
Charge
heterogeneity
analysis,
a
Critical
Quality
Attribute
these
biomolecules
used
Control,
is
often
performed
using
separative
analytical
techniques
such
as
imaged
capillary
Isoelectric
Focusing
(icIEF).
Recognized
gold
standard
by
industry,
icIEF
leverages
pH
gradient
provide
high-resolution
profiling
charge
variants
biotherapeutics.
In
this
review,
critical
experimental
parameters
for
method
development
context
biotherapeutic
drug
QC
will
be
discussed.
Key
aspects,
including
sample
preparation,
properties,
carrier
ampholytes,
stabilizers,
detection
examined,
supported
recent
literature.
Advances
technology
its
expanding
applications
underline
robustness,
reproducibility,
compliance
with
regulatory
standards,
affirming
pivotal
role
ensuring
identity
consistency
biological
products.
