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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 |
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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| 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
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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
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In suspension at
neutral pH;
30 min at 121 °C |
| Supplied
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Suspension in 20
% Ethanol |
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Flow Properties
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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Purification of Rabies Virus
The example in Figure 3 illustrates the high degree of concentration,
purification and yields obtained with Cellufine Sulfate on typical
viral preparations.
| 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.
| 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 :
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20x |
| Concentration Factor : |
28x (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.
| 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 |
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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
| 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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| 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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