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Cellufine - Affinity - ET Clean |
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Affinity Chromatography Media for Endotoxin |
Removal Cellufine |
TM |
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ET Clean |
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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.
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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°.
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Electron micrograph of Cellufine™ ETclean-S beads. |
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Partial Structure |
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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 |
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*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.
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Selective adsorption of endotoxin (LPS) from a bovine serum albumin (BSA) solution by Cellufine ETclean beads. |
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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 ).
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Selective removal of endotoxin from a protein solution by Cellufine™ ETclean beads. |
Sample
Solution |
Cellufine™ ETclean-S |
Cellufine™ ETclean-L |
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| Compound |
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pI |
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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
(%) |
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28,000 |
81 |
99 |
<10 |
95 |
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32,000 |
45 |
99 |
<10 |
97 |
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4,500 |
18 |
99 |
<10 |
98 |
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-globulin |
7.4 |
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5,600 |
20 |
99 |
<10 |
97 |
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1,500 |
15 |
99 |
<10 |
98 |
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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
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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
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Insulin chain A / ETclean-L
- Injection sample (1ml) : 13 mg / ml, 309 EU / ml
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Tranceferrin / ETclean L
- Injection sample (1ml) : 13 mg / ml, 2982 EU / ml
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Ordering Information |
Type |
Quantity |
Catalogue No. |
| Cellufine™ ETclean-S |
10ml |
681984324 |
| Cellufine™ ETclean-L |
10ml |
682985324 |
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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.
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Contact us |
| Americas |
Ivars Bemberis
Director, Cellufine Chromatography
Mobile : (804) 683-7867
Fax : (804) 739-7422
E-mail : ivars10@comcast.net
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| Europe, Asia, and others |
Emil Zavadsky, Ph.D.
Director, Cellufine Chromatography
Mobile : +41/79-206 81 60
Fax : +41/24-499 30 49
E-mail : emil.zavadsky@bluewin.ch
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Ordering Information |
Chisso Corporation
Fine Chemicals Sales Dept.
2-1, Otemachi 2-Chome, Chiyoda-ku,
Tokyo 100-8105 Japan
Tel : +81-3-3243-6150
Fax : +81-3-3243-6219
E-mail : cellufine@chisso.co.jp
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Chisso America Incorporated
555 Theodore Fremd Ave.,
Suite C-206,
Rye, New York 10580 U.S.A.
Tel : 914-921-5400
Fax : 914-921-8822
E-mail : Cellufine@chissony.com
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