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Cellufine Sulfate
Cellufine ET clean
Cellufine PB
Cellufine Phosphate
Cellufine Amino en Formyl
Cellufine Phenyl en Butyl
Cellufine A,Q en C
Cellufine GCL-2000
Cellufine GH-25
Cellufine Mini-Column

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Cellufine Phenyl en Butyl
For purification of proteins and macromolecules.

Hydrophobic Interaction Chromatography (HIC) is a method which separates proteins on the basis of their differential interactions with a mildly hydrophobic surface.

HIC media are porous chromatography particles, manufactured from crosslinked cellulose to which either a butyl, phenyl or octyl functionality has been covalently bonded via a short spacer.

Factors which affect hydrophobic interactions include: salt concentrations, temperature, pH, surfactant and organic solvents. Usually the higher the ionic strength (salt concentration) the stronger the hydrophobic bond. Consequently the interaction is enhanced by conditions inverse to that of ion exchange chromatography. HIC is, therefore, an effective complementary tool for separating and purifying substances which are difficult or cannot be separated by ion exchange.

Partial Structure:

Butyl, Phenyl & Octyl


Features:

• Spherical particles exhibiting high mechanical strength
• Butyl, Phenyl, Octyl functionality
• Pre-swollen
• Virtually no shrinkage or swelling
• Stable in organic solvents and surfactants
• Stable coupling chemistry
• Resistant to 0.2 M NaOH
• Autoclavable (121 °C, 20 min)


Benefits:

• High flow rates allowing rapid chromatography and direct scale-up
• Enables optimum selectivity to be obtained
• Easy packing
• Easy large scale operation. No shrinkage at high salt concentrations
• Enables range of solvent systems to be utilized
• Resistant to cleaning and elution conditions
• Sterilizable
• Regulatory support

Hydrophobicity of Matrix:

The degree of hydrophobicity increases in the order of Butyl < Phenyl < Octyl. In general hydrophobic proteins will be more strongly adsorbed to Cellufine Octyl than Cellufine Butyl. However, if the protein is too strongly adsorbed, difficulty may be experienced in elution. The aromatic nature of the Cellufine Phenyl may, in certain cases, give improved selectivity compared to either the Butyl or Octyl matrices. Consequently it is difficult to generalize and each application needs to be evaluated separately to select the optimal media functionality.

 Figure1

Column : 8.2 x 150 mm
Column Vol. : 8 ml
Buffer : 2.0 – 0.0M
Ammonium Sulfate
in 0.01M phosphate, pH 7.0
Flow Rate : 1.32 ml/min
Sample : 5 mg/3 ml– 100 µl

 

 

 

 


Figure 1. The retention increases with an increase in the carbon chain and aromatic structure of the functional group as a result of stronger hydrophobic interaction.

Flow Properties:
The semi-rigid structure of Cellufine HIC, combined with the spherical bead shape, gives excellent flow rates with higher operating pressures. Flow rates in excess of 100 cm/hr are achieved at pressure drops of 1 bar, even in large diameter process columns.

Figure2

 

 Buffer : 0.01M Phosphate buffer (pH 7.0)
 Temperature : 23 °C
 Columns : (a) 22 x 300 mm
 (Vc = 0.11 liters)
 (b) 90 x 200 mm
 (Vc = 1.27 liters)
 (c) 250 x 250 mm
 (Vc = 12.26 liters)

 

 

 

 

Adsorption Capacity and Recovery Ratio:
 

 

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