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Overexpression, Purification and Validation of Antigenic Salmonella enterica Serovar Typhi Proteins Identified from LC-MS/MS.
[typhoid]
In
our
earlier
study
,
an
immunoblot
analysis
using
sera
from
febrile
patients
revealed
that
a
50
-
kDa
band
from
an
outer
membrane
protein
fraction
of
Salmonella
enterica
serovar
Typhi
was
specifically
recognized
only
by
typhoid
sera
and
not
sera
from
other
febrile
illnesses
.
Here
,
we
investigated
the
identities
of
the
proteins
contained
in
the
immunogenic
50
-
kDa
band
to
pinpoint
antigens
responsible
for
its
immunogenicity
.
We
first
used
LC
-
MS
/
MS
for
protein
identification
,
then
used
the
online
tool
ANTIGENpro
for
antigenicity
prediction
and
produced
recombinant
proteins
of
the
lead
antigens
for
validation
in
an
enzyme-linked
immunosorbent
assay
(
ELISA
)
.
We
found
that
proteins
TolC
,
GlpK
and
SucB
were
specific
to
typhoid
sera
but
react
to
antibodies
differently
under
native
and
denatured
conditions
.
This
difference
suggests
the
presence
of
linear
and
conformational
epitopes
on
these
proteins
.
Diseases
Validation
Diseases presenting
"produced recombinant proteins of the lead antigens"
symptom
typhoid
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