Cellufine® Sulfate

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

NameDatasheetPacksize Order
Cellufine Sulfate Affinity Chromatography Media10 lt View
Cellufine Sulfate Affinity Chromatography Media-10 ml View
Cellufine Sulfate Affinity Chromatography Media5 lt View
Cellufine Sulfate Affinity Chromatography Media50 ml View
Cellufine Sulfate Affinity Chromatography Media500 ml View
MC Cellufine Sulfate1 x 5 ml View
MC Cellufine Sulfate5 x 1 ml View
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:
Hepatitis B Surface Antigen:

 

>3 mg/ml
7 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

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