Electrophoresis,
Journal Year:
2024,
Volume and Issue:
unknown
Published: April 5, 2024
Abstract
Capillary
electrophoresis
(CE)
interfaced
to
mass
spectrometry
(MS)
with
electrospray
ionization
typically
incorporates
acidic
additives
or
organic
solvents
assist
in
ionization.
Vibrating
sharp‐edge
spray
(VSSI)
is
a
voltage‐free
method
interface
CE
and
MS
that
does
not
require
these
additives,
making
it
appealing
for
protein
analyses.
CE–VSSI
nanoflow
sheath
separations
are
performed
low
ionic
strength
aqueous
solutions
the
reduce
suppression.
Serine
also
included
analyte
adduction.
Proteins
detected
2.5–10
µM
range,
corresponding
an
injected
range
of
0.1–1.2
ng.
The
anionic
proteins
β‐lactoglobulin
transferrin
resolved
using
unmodified
fused
silica
capillary
because
they
do
exhibit
nonspecific
surface
adsorption.
Conversely,
cationic
cytochrome
c
,
ribonuclease
A,
α‐chymotrypsinogen
A
background
electrolytes
overcome
Alternatively,
semipermanent
coating
comprised
self‐assembled
lipids
overcomes
adsorption
at
neutral
pH.
Separations
zwitterionic
hybrid
coatings
complete
within
15
6
min,
respectively.
dimeric
form
triosephosphate
isomerase
was
observed
60
µM,
19
ng,
by
dropping
temperature
inlet.
Over
the
past
decades,
number
of
therapeutic
protein
pipelines
in
early-phase
clinical
studies
has
increased
dramatically.
The
rapid
growth
pipeline
underscores
need
to
accelerate
early-stage
development
and
enable
fast
first-in-human
(FIH)
trials
bring
novel
therapies
patients.
Across
industry,
various
efforts
have
been
developed
achieve
this
goal.
In
communication,
a
platform
analytical
method
validation
approach
used
by
MSD
for
FIH
programs
is
described.
A
case
study
from
release
panel,
polysorbate
80
(PS-80)
utilized
illustrate
workflow.
approach,
historical
data
within
same
modality
are
summarized
supplemented
with
statistical
analyses
justify
limited
future
projects.
strategy
reduced
overall
timeline
up
4
months
1-2
successfully
implemented
filings.
This
communication
provides
insights
pharmaceutical
companies
interested
developing
approaches
studies.
Electrophoresis,
Journal Year:
2023,
Volume and Issue:
44(15-16), P. 1247 - 1257
Published: April 20, 2023
Capillary
zone
electrophoresis
ultraviolet
(CZE-UV)
has
become
increasingly
popular
for
the
charge
heterogeneity
determination
of
mAbs
and
vaccines.
The
ε-aminocaproic
acid
(eACA)
CZE-UV
method
been
used
as
a
rapid
platform
method.
However,
in
last
years,
several
issues
have
observed,
example,
loss
electrophoretic
resolution
or
baseline
drifts.
Evaluating
role
eACA
on
reported
issues,
various
laboratories
were
requested
to
provide
their
routinely
methods,
background
electrolyte
compositions.
Although
every
laboratory
claimed
use
He
et
al.
method,
most
methods
actually
deviate
from
He's.
Subsequently,
detailed
interlaboratory
study
was
designed
wherein
two
commercially
available
(Waters'
Mass
Check
Standard
mAb
[pI
7]
NISTmAb
9])
provided
each
laboratory,
along
with
protocols
short-end,
high-speed,
long-end,
high-resolution
Ten
participated
using
own
instruments,
commodities,
showing
excellence
performance
(relative
standard
deviations
[RSDs]
percent
time-corrected
main
peak
areas
0.2%
1.9%,
RSDs
migration
times
0.7%
1.8%
[n
=
50
per
laboratory],
analysis
some
cases
short
2.5
min).
This
clarified
that
is
not
reason
abovementioned
variations.
Electrophoresis,
Journal Year:
2024,
Volume and Issue:
unknown
Published: April 5, 2024
Abstract
Capillary
electrophoresis
(CE)
interfaced
to
mass
spectrometry
(MS)
with
electrospray
ionization
typically
incorporates
acidic
additives
or
organic
solvents
assist
in
ionization.
Vibrating
sharp‐edge
spray
(VSSI)
is
a
voltage‐free
method
interface
CE
and
MS
that
does
not
require
these
additives,
making
it
appealing
for
protein
analyses.
CE–VSSI
nanoflow
sheath
separations
are
performed
low
ionic
strength
aqueous
solutions
the
reduce
suppression.
Serine
also
included
analyte
adduction.
Proteins
detected
2.5–10
µM
range,
corresponding
an
injected
range
of
0.1–1.2
ng.
The
anionic
proteins
β‐lactoglobulin
transferrin
resolved
using
unmodified
fused
silica
capillary
because
they
do
exhibit
nonspecific
surface
adsorption.
Conversely,
cationic
cytochrome
c
,
ribonuclease
A,
α‐chymotrypsinogen
A
background
electrolytes
overcome
Alternatively,
semipermanent
coating
comprised
self‐assembled
lipids
overcomes
adsorption
at
neutral
pH.
Separations
zwitterionic
hybrid
coatings
complete
within
15
6
min,
respectively.
dimeric
form
triosephosphate
isomerase
was
observed
60
µM,
19
ng,
by
dropping
temperature
inlet.
