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Cellufine - Affinity - Sulfate |
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| NEW!! Cellufine Mini-Column
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Affinity Chromatography Media Cellufine |
TM |
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Sulfate |
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For concentration, purification and depyrogenation of virus, viral/microbial antigens and heparin-binding proteins |
Advances in vaccines and clinical diagnostics have created an increasing demand for large volumes of highly purified and concentrated virus and viral or microbial antigens. Cellufine Sulfate affinity media is a simple, rapid and effective means for concentration, purification and depyrogenation of these important products.
Cellufine Sulfate eliminates cumbersome, time-consuming and potentially unsafe classical ultra-centrifugation and density gradient methods. It can also provide a significant improvement in concentration and purity. Cellufine Sulfate can reduce or eliminate the expense, ligand leakage and reproducibility problems associated with immobilized dextran sulfate, chondroitin sulfate or heparin.
Elution of the bound product is affected through simple stepwise or gradient increases in ionic strength.
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Features |
• Affinity for a wide range of live, killed or disrupted viruses, viral or microbial antigens and heparin-binding proteins
• Closed column operation assures safety and product sterility
• Endotoxins do not bind, allowing a rapid and contaminant free depyrogenation
• Rigid, high-strength beads
• Autoclavable
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Benefits |
• More effective than ultracentrifugation at removing contaminants from culture media and host cells
• Avoids excessive product handling and safety concerns, particularly with viral preparations
• Simultaneous concentration and purification improve yield, reduce processing steps, time and costs
• Gentle binding and elution conditions provide high capacity and product yield
• Resists compression, providing rapid flow for high-speed processing, even in large columns, making it easily scalable
• Resistant to chemical depyrogenation with base and chemically sterilizable with formalin
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Characteristics |
| Support Matrix |
Cellulose |
| Particle Size |
44 – 105 µm |
| Particle Shape |
Spherical |
| Gel Exclusion Limit |
3kD |
| Activated Group |
Sulfate Ester |
| Total Sulfur |
700 µg/g dry |
Protein Binding Capacity
Lysozyme :
Hepatitis B Surface
Antigen
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3 mg/ml
7 mg/ml |
| Environmental Resistance |
Resistant to 0.1M NaOH,
0.1 % of 37 % Formalin |
| Operating Pressure |
<2 bar (30 psi) |
| Autoclavable |
In suspension at neutral pH;
30 min at 121 °C |
| Supplied |
Suspension in 20 %
Ethanol |
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Flow Properties |
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The nearly rigid properties of the spherical cellulose support matrix allow
outstanding flow properties, particularly in large production columns.
Column A: 90 x 200 mm
Column B: 350 x 200 mm
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Virus, Viral/Microbial Antigens |
Viruses |
Viral/Microbial Agents |
• Rabies*
• Influenza*
• Japanese Enchephalitis*
• Feline Leukemia
• Feline Herpes
• Feline Calicivirus
• Respiratory Syncytial Virus
• Human Herpes Simplex
• Human Measles
• Human Parainfluenza |
• Herpes Simplex gA and gB
Glycoprotein Subunits*
• Hepatitis B Surface Antigen
• Filamentous Hemagglutinin from
B. pertussis *
• Leucocytosis Promoting Factor
Hemagglutinin* |
| *These applications are covered by US and foreign process patents.
Please inquire regarding details and licensing arrangements. |
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Virus, Viral/Microbial Antigens |
The example in Figure 3 illustrates the high degree of concentration, purification and yields obtained with Cellufine Sulfate on typical viral preparations. |
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Column : 50 x 70 mm (140 ml)
Buffer : 0.01M Phosphate (pH 7.2)
Eluant : 1M NaCl/0.01M Phosphate (pH 7.2)
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Load |
Eluate |
| Volume (ml) |
4,200 |
50 |
| Virus titer |
32 |
4,096 |
| Protein (µg/ml) |
8.5 |
14 |
| Yield (%) |
100 |
152 |
| Purification factor |
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79x |
| Concentration factor |
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126x |
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Purification of Influenza Virus |
Hen’s egg allantoic fluid was loaded directly onto a 33.3 mL gel bed and 94.5%virus was recovered in the eluate fraction. |
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Volume (ml) |
Virus Titer |
TCA-N µg/ml |
Recovery (%) |
Fold Purification |
| Allantoic Fluid |
4200 |
77 |
337.1 |
100 |
1 |
| Wash |
6700 |
1 |
209.2 |
2.1 |
- |
| Eluate |
170 |
1797 |
448.0 |
94.5 |
20.1 |
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Column : 50 x 170 mm
Buffer : 0.01M Phosphate pH 7.4
Wash : 0.01M Phosphate pH 7.2 + 0.2M NaCl
Elution : 0.01M Phosphate pH 7.0 + 1.5M NaCl
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Antigenic Protein Purification and Depyrogenation |
Cellufine Sulfate is ideal for depyrogenating virus and other microbial extracts because it does not bind endotoxins. Figure 4 shows the purification of filamentous hemagglutinin (FHA) from the whooping cough bacterium Bordetella pertussis.
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| Column : |
16 x 70 mm (20 ml) |
| Sample : |
800 ml B. pertussis culture fluid
(endotoxin titer > 1015 by Limulus lysate test) |
| Buffer : |
0.01M Phosphate (pH 7.6) |
| Eluant : |
1M NaCl/0.01M Phosphate (pH 7.6) |
| FHA Yield : |
94% |
| Purification Factor : |
20x |
| Concentration Factor : |
128x (30 ml product) |
| Endotoxin : |
Below standard level by Limulus lysate, rabbit pyrogen and mouse toxicity tests |
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Protein Purification |
Cellufine Sulfate mimics the affinity of heparin or dextran sulfate for many proteins. It can function as an affinity support for selected plasma proteins, cellular growth factors and lipases. Its capacity comparable to conventional heparin gels.
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Binding Proteins |
Non-binding Proteins |
• Antithrombin III
• β-Lipoprotein
• Complement C5, C6, C8
• Complement C3 Activator
• Tryspin
• Tryspin Inhibitor
• Chymotrypsinogen
• Lysozyme
• Urease
• Catalase
• Factor IX |
• Albumin
• α-Lipoprotein
• Complement C3, C9
• Complement C1, C3b
Inactivators
• IgG
• Ceruloplasmin
• α2-Macroglobulin
• RNase
• Bacitracin
• Glucose Oxidase |
| *Binding and elution are extremely rapid and very fine separations can be generated in gradient mode. |
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Purification of Partially Purified Casein Kinase II from Calf Thymus |
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| Column : |
10 x 20 mm |
| Sample : |
7 ml |
| Buffer : |
50mM Tris-HCl (pH 7.9)
+ 50mM MgCl2
+ 0.1mM EDTA
+ 0.1mM PMS
+ 0.5mM DTT
+ 25 % glycerol |
| Eluant : |
0.05 – 1.0M NaCl in buffer |
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Ordering Information |
| Description |
Quantity |
Catalogue No |
| Cellufine Sulfate |
10 ml |
676943324 |
| 50 ml |
19845 |
| 500 ml |
19846 |
| 5 Liters |
19847 |
| 10 Liters |
19849 |
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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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