Cellufine® Sulfate Affinity Media
Affinity Chromatography Media
For use in the 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 based on spherical cellulose beads functionalized with a low concentration of sulfate esters, and is a simple, rapid and effective means for the concentration, purification and depyrogenation or viral and microbial antigens and specific proteins.
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
- Can reduce or eliminate the expense, ligand leakage and reproducibility problems associated with immobilized dextran sulfate, chondroitin sulfate or heparin
| Cellufine Sulfate Characteristics | |
|---|---|
| Support Matrix | Cellulose |
| Particle Size | ca. 40-120µ |
| Particle Shape | Spherical |
| Gel Exclusion Limit | ca. 3kD |
| Activated Group | Sulfate Ester |
| Total Sulfur | >700 ug/g dry |
|
Protein Binding Capacity
Lysozyme: |
>3 mg/ml |
| Environmental Resistance | Resistant to 0.1M NaOH, 0.1% of 37% Formalin |
| Operating Pressure | <2 bar (30 psi) in suspension at neutral pH |
| Autoclavable | In suspension a neutral pH; 30 min at 121°C |
| Supplied | Suspension in 20% Ethanol |
Cellufine resin was developed by JNC and is available from AMSBIO in EU and the US.
Flow Properties
The nearly rigid properties of the spherical cellulose support matrix allow outstanding flow properties, particularly in large production columns.
Virus and Viral/Microbial Antigens
There are many applications of Cellufine Sulfate in the concentration or purification of viral and microbial agents, proteins and viruses.
| Viruses | Viral/Microbial Agents |
|---|---|
| Rabies* Influenza* Japanese Encephalitis* 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* Leukocytosis Promoting Factor Hemagglutinin* |
*These applications are covered by US and foreign process patents. Please inquire regarding details and licensing arrangements.
Purification Of Rabies Virus From Chick Embryo Tissue Culture Fluid
This example illustrates the high degree of concentration, purification, and yields obtained with Cellufine Sulfate on typical viral preparations:
Concentration and Purification of Virus with Cellufine Sulfate
| Load | Elute | |
|---|---|---|
| Volume (mL) | 4,200 | 50 |
| Virus Titer | 32 | 4,096 |
| Protein (µg/mL) | 8.5 | 14 |
| Yield (%) | 100 | 152 |
| Purification Factor | - | 79x |
| Concentration Factor | - | 126x |
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
| 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 |
Purification of Influenza virus from hen’s egg allantoic fluid. Column: 50 x 170mm, 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
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, and its capacity is comparable to conventional heparin gels.
| Binding Proteins | Non-binding Proteins |
|---|---|
| Antihrombin III | Albumin |
| β-Lipoprotein | α-Lipoprotein |
| Complement C5, C6, C8 | Complement C3, C9 |
| Complement C3 Activator | Complement C1, C3b Inactivators |
| Tryspin | IgG |
| Tryspin Inhibitor | Ceruloplasmin |
| Chymotrypsinogen | α2-Macroglobulin |
| Lsozyme | RNase |
| Urease | Bacitracin |
| Catalase | Glucose Oxidase |
| Factor IX |
*Binding and elution are extremely rapid and very fine separations can be generated in gradient mode
Purification of Partially Purified Casein Kinase II from Calf Thymus
Cellufine™ is the trademark of JNC Corporation, Tokyo, Japan
