JNC CORPORATION : Cellufine : Products

Affinity Chromatography Media
for Endotoxin Removal
Cellufine™ ET clean

Cellufine ET clean S > Reference > MSDS / Technical Information > Catalog No.


Cellufine ET clean L > Reference > MSDS / Technical Information > Catalog No.



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Introduction

 

The CellufineETclean 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 JNC Corporation.


The CellufineETclean 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

The CellufineETclean 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(TM) ETclean-S beads.
  Electron micrograph of
Cellufine™ ETclean-S beads.


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Partial Structure
Partial Structure


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Characteristics

Name Supplied Wet Bead
Diameter
Pore Size*
Cellufine™ ETclean-S a slurry in 20 % ethanol ca. 40-130 μm Mlim 2000
Cellufine™ ETclean-L a slurry in 20 % ethanol ca. 40-130 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.


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Selective adsorption of endotoxin (LPS) from a bovine serum albumin (BSA) solution by Cellufine ETclean beads.

ETclean

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

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
(%)
Ovalbumin 4.6
28,000 81 99 <10 95
BSA 4.9
32,000 45 99 <10 97
Myoglobin 6.8
4,500 18 99 <10 98
γ-globulin 7.4
5,600 20 99 <10 97
Cytochrome C 10.6
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.


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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

Application Data

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

Application Data

Insulin chain A / ETclean-L
- Injection sample (1ml) : 13 mg / ml, 309 EU / ml

Application Data

Tranceferrin / ETclean L
- Injection sample (1ml) : 13 mg / ml, 2982 EU / ml

Application Data


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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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