Protein-Centric Analysis of Personalized Antibody Repertoires Using LC-MS-Based Fab-Profiling on a timsTOF
Journal of the American Society for Mass Spectrometry,
Journal Year:
2024,
Volume and Issue:
35(6), P. 1292 - 1300
Published: April 25, 2024
Endogenous
antibodies,
or
immunoglobulins
(Igs),
abundantly
present
in
body
fluids,
represent
some
of
the
most
challenging
samples
to
analyze,
largely
due
immense
variability
their
sequences
and
concentrations.
It
has
been
estimated
that
our
can
produce
billions
different
Ig
proteins
with
isotypes,
making
individual
analysis
seemingly
impossible.
However,
recent
advances
protein-centric
proteomics
using
LC-MS
coupled
Orbitrap
mass
analyzers
profile
intact
Fab
fragments
formed
by
selective
cleavage
at
IgG-hinge
revealed
IgG
repertoires
may
be
less
diverse,
albeit
unique
for
each
donor.
Serum
seem
dominated
a
few
hundred
clones
cumulatively
make
up
50–95%
total
content.
Enabling
such
analyses
required
careful
optimization
chromatography
analysis,
as
all
analytes
are
highly
alike
(46–51
kDa)
sequence.
To
extend
opportunities
this
mass-spectrometry-based
profiling
antibody
repertoires,
we
here
report
evaluation
an
alternative
MS
platform,
namely,
timsTOF,
repertoire
profiling.
The
timsTOF
analyzer
gained
traction
years
peptide-centric
found
wide
applicability
plasma
proteomics,
affinity
HLA
peptidomics,
name
few.
platform
explored.
Here,
demonstrate
adapted
perform
LC-MS-based
repertoires.
In
side-by-side
comparison
extracted
serum
qualitatively
quantitatively,
whereby
particular
sensitivity
excels.
Future
incorporation
advanced
top-down
capabilities
on
very
valuable
thus
also
personalized
Language: Английский
CE-MS and CE-MS/MS for the Multiattribute Analysis of Monoclonal Antibody Variants at the Subunit Level
Journal of Pharmaceutical and Biomedical Analysis,
Journal Year:
2024,
Volume and Issue:
252, P. 116495 - 116495
Published: Oct. 2, 2024
Language: Английский
Flow Rate Determination of the Nanoflow Sheath Liquid CE–MS‐Coupling Applying the nanoCEasy Interface
Electrophoresis,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 28, 2024
ABSTRACT
In
recent
years,
nanoflow
sheath
liquid
(nanoSL)
interfaces
have
been
used
more
commonly
for
capillary
electrophoresis–mass
spectrometry
(CE–MS)
coupling
due
to
their
high
sensitivity
combined
with
flexibility
in
CE
separation
conditions.
So
far,
the
exact
amount
of
(SL)
and,
thus,
dilution
effect
remain
unknown
self‐supplying
type
nanoSL
interfaces.
To
quantify
SL
flow
rates,
we
present
here
an
approach
determination
rate
based
on
isotopically
labeled
standards
using
nanoCEasy
interface.
and
pumped
are
spiked
atrazine
d5‐atrazine,
respectively,
enabling
total
different
electrospray
(ES)
Generally,
rates
increase
higher
ES
voltages.
Only
at
voltages,
larger
emitter
sizes
lead
rates;
low
independent
size.
Additionally,
smaller
can
generally
be
operated
lower
Visualizing
ES,
a
spray
angle.
Higher
may
decrease
signal
intensity
by
diluting
CE‐separated
analytes.
Indeed,
this
is
observed
flow‐injected
analytes
CE–MS
analysis
amino
acid
standard
mix
under
aqueous
acidic
Therefore,
preferred
electroosmotic
(EOF)
applications.
However,
EOF
applications,
voltages
required
prevent
accumulation
background
electrolyte
(BGE)
tip,
which
otherwise
leads
peak
broadening
inefficient
ionization.
Overall,
presented
data
enable
selection
stable
sensitive
settings
Language: Английский