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Production of scFv-conjugated affinity silk film and its application to a novel enzyme-linked immunosorbent assay.
[wiskott-aldrich syndrome]
Bombyx
mori
(
silkworm
)
silk
proteins
have
been
utilized
as
unique
biomaterials
for
various
medical
applications
.
To
develop
a
novel
affinity
silk
material
,
we
generated
a
transgenic
silkworm
that
spins
silk
protein
containing
the
fibroin
L-
chain
linked
with
the
single
-chain
variable
fragment
(
scFv
)
as
a
fusion
protein
.
Previously
,
the
scFv-conjugated
"
affinity
"
silk
powder
specifically
immunoprecipitated
its
target
protein
,
Wiskott-
Aldrich
syndrome
protein
.
To
expand
the
applicability
of
affinity
silk
materials
,
we
processed
the
scFv-conjugated
silk
protein
into
a
thin
film
by
dissolving
it
in
lithium
bromide
,
then
drying
it
in
the
wells
of
96
-
well
plates
.
Enzyme-linked
immunosorbent
assay
demonstrated
specific
detection
of
Wiskott-
Aldrich
syndrome
protein
,
both
as
a
recombinant
protein
and
in
its
native
form
extracted
from
mouse
macrophages
.
These
findings
suggest
that
this
scFv-conjugated
silk
film
serves
as
the
basis
for
an
alternative
immunodetection
system
.
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"been utilized as unique biomaterials"
symptom
wiskott-aldrich syndrome
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