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Oligosaccharide analysis in urine by maldi-tof mass spectrometry for the diagnosis of lysosomal storage diseases.
[gm1 gangliosidosis]
There
are
45
known
genetic
diseases
that
impair
the
lysosomal
degradation
of
macromolecules
.
The
loss
of
a
single
lysosomal
hydrolase
leads
to
the
accumulation
of
its
undegraded
substrates
in
tissues
and
increases
of
related
glycoconjugates
in
urine
,
some
of
which
can
be
detected
by
screening
of
free
oligosaccharides
(
FOS
)
in
urine
.
Traditional
1
-
dimensional
TLC
for
urine
oligosaccharide
analysis
has
limited
analytical
specificity
and
sensitivity
.
We
developed
fast
and
robust
urinary
FOS
and
glycoaminoacid
analyses
by
MALDI-time-of-flight
/
time-of-flight
(
MALDI-TOF
/
TOF
)
mass
spectrometry
for
the
diagnosis
of
oligosaccharidoses
and
other
lysosomal
storage
diseases
.
The
FOS
in
urine
equivalent
to
0
.
09
mg
creatinine
were
purified
through
sequential
passage
over
a
Sep
-
Pak
C
18
column
and
a
carbograph
column
and
were
then
permethylated
.
MALDI-TOF
/
TOF
was
used
to
analyze
the
permethylated
FOS
.
We
studied
urine
samples
from
individuals
in
7
different
age
groups
ranging
from
0
-
1
months
to
≥
17
years
as
well
as
urine
from
known
patients
with
different
lysosomal
storage
diseases
.
We
identified
diagnostic
urinary
FOS
patterns
for
α-mannosidosis
,
galactosialidosis
,
mucolipidosis
type
II
/
III
,
sialidosis
,
α-
fucosidosis
,
aspartylglucosaminuria
(
AGU
)
,
Pompe
disease
,
Gaucher
disease
,
and
GM
1
and
GM
2
gangliosidosis
.
Interestingly
,
the
increase
in
urinary
FOS
characteristic
of
lysosomal
storage
diseases
relative
to
normal
FOS
appeared
to
correlate
with
the
disease
severity
.
The
analysis
of
urinary
FOS
by
MALDI-TOF
/
TOF
is
a
powerful
tool
for
first
-tier
screening
of
oligosaccharidoses
and
lysosomal
storage
diseases
.
Diseases
Validation
Diseases presenting
"some of which can be detected by screening of free oligosaccharides"
symptom
gm1 gangliosidosis
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