A Robust and Automated Platform for Charge Detection Mass Spectrometry of Megadalton Biotherapeutics
Analytical Chemistry,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 19, 2025
Gene
therapies
based
on
adeno-associated
viruses
are
an
emerging
area
with
high
potential
to
improve
human
health.
Current
quality
control
techniques
assess
contaminates
and
byproducts
from
the
virus
(AAV)
production
pipelines
lacking
in
robustness
throughput.
To
address
these
limitations,
we
coupled
automated
microfluidic
device
called
SampleStream
Orbitrap-based
charge
detection
mass
spectrometry
(SS-CDMS).
We
demonstrate
that
SS-CDMS
workflow
performs
AAV
analysis
under
15
min
per
sample
a
completely
autonomous
manner.
The
enables
rapid
assessment
of
key
attributes
(CQAs),
such
as
molecular
weight
content
ratio
formulations
small
requirement
(<2
×
109
capsids)
without
being
limited
by
concentration.
Additionally,
this
work
shows
for
be
implemented
at
various
stages
pipeline
through
effective
clean
up
more
complex
matrices
cell
culture
media.
Language: Английский
Roughhousing with Ions: Surface-Induced Dissociation and Electron Capture Dissociation as Diagnostics of Q-Cyclic IMS-TOF Instrument Tuning Gentleness
Journal of the American Society for Mass Spectrometry,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 7, 2024
Native
mass
spectrometry
can
characterize
a
range
of
biomolecular
features
pertinent
to
structural
biology,
including
intact
mass,
stoichiometry,
ligand-bound
states,
and
topology.
However,
when
an
instrument's
ionization
source
is
tuned
maximize
signal
intensity
or
adduct
removal,
it
possible
that
the
complex's
tertiary
quaternary
structures
be
rearranged
in
way
no
longer
reflect
its
native-like
conformation.
This
could
affect
downstream
ion
activation
experiments,
leading
erroneous
conclusions
about
structure.
One
strategy
surface-induced
dissociation
(SID),
which
generally
causes
protein
complexes
dissociate
along
weakest
subunit
interfaces,
revealing
critical
information
topology
connectivity.
If
structure
has
been
disturbed,
then
SID
fingerprint
will
shift
as
well.
Thus,
was
used
diagnose
source-induced
rearrangement
help
tune
other
upstream
transmission
regions
strike
balance
between
intensity,
conserving
Complementary
SID,
electron-capture
(ECD)
also
after
in-source
confirm
interfaces
were
rearranged,
opening
electron
capture
subsequent
dissociation.
These
results
provide
valuable
guide
for
new
practitioners
native
highlight
importance
using
standard
tuning
instrument
platforms
optimal
performance.
Language: Английский
Infrared photoactivation enables nano-DESI MS of protein complexes from tissue on a linear ion trap mass spectrometer.
Published: Aug. 9, 2024
Native
mass
spectrometry
analysis
of
proteins
directly
from
tissues
can
be
performed
using
nanospray-desorption
electrospray
ionization
(nano-DESI).
Typically,
supplementary
collisional
activation
is
essential
to
decluster
protein
complex
ions
solvent,
salt,
detergent
and
lipid
clusters
that
comprise
the
ion
beam.
As
an
alternative,
we
have
implemented
declustering
by
infrared
(IR)
photoactivation
on
a
linear
trap
spectrometer
equipped
with
CO2
laser.
The
prototype
system
demonstrates
intact
up
approx.
50
kDa
in
molecular
weight
were
sampled
brain
eye
lens
nano-DESI.
For
example,
signals
attributable
different
metal
binding
states
hSOD1G93A
homodimers
(approx.
32
kDa)
separated
only
6
Th
(10+
ions)
resolved
IR
declustering,
but
not
activation.
We
found
outperform
its
ability
reduce
chemical
background
non-specific
nano-DESI
also
situ
native
MS
low-cost
potential
spectrometers
for
this
type
analysis.
Language: Английский
Infrared Photoactivation Enables nano-DESI MS of Protein Complexes in Tissue on a Linear Ion Trap Mass Spectrometer
Journal of the American Society for Mass Spectrometry,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 27, 2024
Native
mass
spectrometry
analysis
of
proteins
directly
from
tissues
can
be
performed
by
using
nanospray-desorption
electrospray
ionization
(nano-DESI).
Typically,
supplementary
collisional
activation
is
essential
to
decluster
protein
complex
ions
solvent,
salt,
detergent,
and
lipid
clusters
that
comprise
the
ion
beam.
As
an
alternative,
we
have
implemented
declustering
infrared
(IR)
photoactivation
on
a
linear
trap
spectrometer
equipped
with
CO
Language: Английский