Introduction
The Cellufine™ ETclean is
poly(ε-lysine)
immobilized Cellufine™ (cellulose
spherical
beads).
The beads
bind
and remove
endotoxin
from
your
sample
solution.
The poly(ε-lysine)
is a
microbial
poly(amino
acid)
that
consist
of 30-35
lysine
residues
produced
by Streptomyces
albulus.
The poly(ε-lysine)
as ligand
and the
cellulose
beads
act as
matrix
ands
are products
of Chisso
Corporation. |
The Cellufine™ ETclean endotoxin
removing
beads
were
developed
jointly
by
Kumamoto
University
and
Chisso.
The
poly(ε-lysine)
was
immobilized
onto
chloromethyloxirane-activated
cellulose
beads.
The
beads
are
a
stable
affinity
beads
that
are
resistant
against
the
cleanup
solutions,
which
include
0.2
M
sodium
hydroxide
and
2
M
sodium
chloride.
The Cellufine™ ETclean can
remove
endotoxin
from
a
cellular
product
solution
at
physiological
pH,
ionic
strength
of μ =
0.02-1.0,
and
0° -25C°. |
 |
| |
Electron
micrograph
of
Cellufine™ ETclean-S
beads. |
|
Partial
Structure
 |
Characteristics
| Name |
Supplied |
Wet
Bead
Diameter |
Pore
Size* |
| Cellufine™ ETclean-S |
a
slurry
in
20
%
ethanol |
45-105 μm |
Mlim 2000 |
| Cellufine™ ETclean-L |
a
slurry
in
20
%
ethanol |
53-125 μm |
>Mlim 2x106 |
|
*The
pore
size
(molecular
weight
exclution;
Mlim)
of the
beads
was estimated
from
calibration
curves
obtained
by size
exclusion
chromatography.
Pullulan
and maltose
were
used
for the
Mlim determination. |
Selective
adsorption
of
endotoxin
(LPS)
from
a
bovine
serum
albumin
(BSA)
solution
by
Cellufine
ETclean
beads.
 Selective
adsorption
of endotoxin
was determined
using
a batchwise
method
with
0.2 g
of the
wet beads
and 2
ml of
a sample
solution
(BSA:
500 μg/ml, E.
coli O111:
B4 LPS:
100 ng/ml,
pH 7.0,
ionic
strength
of μ =
0.05-0.8
). |
Selective
removal
of
endotoxin
from
a
protein
solution
by
Cellufine™ ETclean
beads.
| Sample
Solution |
Cellufine™ ETclean-S |
Cellufine™ ETclean-L |
 |
| Compound |
|
| |
pI |
|
Concentration
of
endotoxin
before
treatment
(pg/ml) |
(μ =
0.05, pH 7.0) |
(μ =
0.05, pH 7.0) |
Concentration
of
endotoxin
after
treatment
(pg/ml) |
Recovery
of
protein
after
treatment
(%) |
Concentration
of
endotoxin
after
treatment
(pg/ml) |
Recovery
of
protein
after
treatment
(%) |
|
|
28,000 |
81 |
99 |
<10 |
95 |
|
|
32,000 |
45 |
99 |
<10 |
97 |
|
|
4,500 |
18 |
99 |
<10 |
98 |
 |
-globulin |
7.4 |
|
5,600 |
20 |
99 |
<10 |
97 |
|
|
1,500 |
15 |
99 |
<10 |
98 |
|
The removal
of endotoxin
was determined
by a
batchwise
method
with
0.3 ml
of wet
adsorbent
and 2
ml of
a protein
solution
(1 mg/ml)
containing
natural
endotoxin. |
Application
Data
ET
removal
Exp.
BSA
/
ETclean
L |
| Column
chromatography |
| - |
Column
size
:
1
X
1.1
cm
(I.D.)
(1.1ml) |
| - |
Flow
rate
0.17
ml
/
min
(10cm
/
h) |
| - |
Buffer
50
mM
PB,
pH
7
+
0.15
mol
NaCl
aq |
| Assay |
| - |
Protein
Abs.
at
280
nm |
| - |
ET
LAL
rate
assay |
| |
-
Injection
sample
(150
ml)
-
BSA
1
mg/ml
ET
100
EU/ml |
| Lysozyme
/
ETclean
L |
| Column
chromatography |
| - |
Column
size
10
x
0.9
cm
(I.D.)
(9.6
ml) |
| - |
Flow
rate
0.5
ml
/
min
(47
cm
/
h) |
| - |
Buffer
1
mM
Tris-HCl,
pH
7.3 |
| - |
Gradient
0 → 1.0
mol
/
l
NaCl
aq. |
| Assay |
| - |
Protein
Abs.
at
280
nm |
| - |
ET
LAL
rate
assay |
| |
| -
Injection
sample
(1ml)
:
14
mg
/
ml |
| Insulin
chain
A
/
ETclean-L |
| -
Injection
sample
(1ml)
:
13
mg
/
ml,
309
EU
/
ml |
| Tranceferrin
/
ETclean
L |
| -
Injection
sample
(1ml)
:
13
mg
/
ml,
2982
EU
/
ml |
 |
Ordering
Information
| Type |
Quantity |
Catalogue
No. |
| Cellufine™ ETclean-S |
10ml |
681984324 |
| Cellufine™ ETclean-L |
10ml |
682985324 |
|
|
References
1) M.
Sakata,
M. Todokoro,
C. Hirayama, American
Biotechnol.
Lab., 20 (2002)
36.
2) M.
Todokoro,
M. Sakata,
S. Matama,
M. Kunitake,
J. Ohkuma,
C. Hirayama, J.
Liq.
Chrom. & Rel.
Technol., 25 (2002)
601.
Cellufine
ET clean
was developped
by Kumamoto
Univ & Chisso
Corp
Joint
Project. |
inquiry
