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Cellufine - Hydrophobic Interaction |
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Hydrophobic Interaction Chromatography Media |
Cellufine |
TM |
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Butyl, Phenyl & Octyl |
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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.
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Partial Structure |
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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)
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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
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Characteristics |
| Support Matrix |
Crosslinked Cellulose |
| Particle Size |
53 – 125 µm |
| Particle Shape |
Spherical beads |
| Exclusion Limit |
4,000 kD |
| Functional Group |
Butyl, Phenyl, Octyl |
| Shrinkage/Swelling |
Negligible |
| pH Stability
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pH 1 – 13 |
| Environmental Resistance |
Resistant to 0.2M NaOH |
| Operating Pressure |
Up to 1 bar (15 psi) |
| Solvent Resistance |
Resistant to detergents, organic solvents, salts |
| Supplied |
Suspension in 20 % Ethanol |
| Density |
1.3 ml/g wet gel |
| Autoclavable |
121 °C, 20 min |
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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.
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| 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 |
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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.
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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
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| 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)
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Adsorption Capacity and Recovery Ratio |
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
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Cellufine Butyl |
Cellufine Phenyl |
Cellufine Octyl |
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Adsorption mg/ml |
Recovery % |
Adsorption mg/mL |
Recovery % |
Adsorption mg/ml |
Recovery % |
| BSA |
25 |
87 |
30 |
92 |
23 |
87 |
| Catalase |
42 |
62 |
35 |
71 |
25 |
62 |
| Myoglobin |
19 |
62 |
11 |
63 |
9 |
61 |
| Glucose Oxidose |
38 |
97 |
37 |
99 |
15 |
100 |
| Ovalbumin |
31 |
87 |
31 |
89 |
19 |
88 |
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| Sample concentration : |
0.1 % |
| Adsorption buffer : |
0.1M phosphate buffer, pH 7.0 + 2M (NH4)2SO4 |
| Recovery buffer : |
0.01M phosphate buffer, pH 7.0 |
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Ordering Information |
| Description |
Quantity* |
Catalogue No |
| Cellufine Butyl |
100 ml
500 ml
5 liters
10 liters |
19905
19906
19879
6769 733 35 |
| Cellufine Phenyl |
100 ml
500 ml
5 liters
10 liters |
19900
19901
19881
6769 753 35 |
| Cellufine Octyl |
100 ml
500 ml
5 liters
10 liters |
19895
19896
19880
6769 743 35 |
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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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