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Cellufine GH-25
For rapid protein desalting buffer exchange and removal of alcohol and detergents.

Cellufine GH-25 desalting media is based on porous, spherical, highly crosslinked cellulose particles. The sharp 3kD exclusion limit allows proteins to pass through the column in the void volume while retarding smaller molecular weight solutes in the internal pores. Outstanding mechanical strength allows operation at high flow rates even in large diameter process scale columns, thus minimizing run times.


Features:

• Mechanical robust spherical particle
• Efficient salt removal
• Hydrophilic
• Pre-swollen
• pH stable 1 – 14 (0.1M HCl, 0.5M NaOH)
• Resistant to organic solvents
• Autoclavable (121 °C, 30 min)


Benefits:

• Enables high flow rates and short run times
• Permits large sample loads (typical: 5 – 30 minute run times in columns from 1 ml to 100 liters, with loads up to 35 % bed volume)
• Low non-specific adsorption, high recovery
• Easy packing
• Easy cleaning and depyrogenation
• Permits use with all commonly used solvents and buffers without shrinkage or swelling
• Sterilizable

Flow Properties:
 

Figure1

 

The specific selection curve of Cellufine GH-25:
 

The specific selection curve of Cellufine GH-25
1: Glycine 75 2: (Gly)2 132 3: (Gly)3 189 4: (Gly)4 246
5: Calcium pantothenate 477 6: Vitamin B12 1355 7: insulin B chain 3495


Applications:

• Desalting before lyophilization or concentration
• Buffer exchanges
• Removal of alcohol or other organic solvents
• Removal of aromatic compounds (e.g., phenol) in purification of nucleic acids
• Removal of detergents used to solubilize proteins (e.g. Triton® X-100, SDS)
• Permits use with all commonly used solvents and buffers without shrinkage or swelling
• Removal of chaotropic agents, (e.g. urea, guanidine)


High Speed Desalting:

Although the rigidity of Cellufine GH-25 makes it ideally suited to large scale column use, its ability to operate at very high flow rates enables the use of smaller columns running multiple cycles giving similar throughput to lower flow rates on larger columns. Unlike conventional chromatography, the performance (as measured by sample load, salt removal, and dilution) of desalting chromatography can actually improve as the flow rate increases, due to decreased sample dilution at high volumetric loads.


Protein Desalting:
 

Figure2

Packing : Cellufine GH-25
Column : 105 x 587 mm Vt = 5086 ml
Mobile Phase : 0.1M NaCl
Flow : 1250 ml/min, 870 ml/h/cm2
Sample : 5 % (w/v BSA) in 1.5M NaCl
Sample Volume : 1272 ml (25 % of column volume)
Protein Recovery : 99.5 %
Salt Exchange : 99 %
Dilution : 1.13x


Alcohol Removal:
 

Fractionation of human blood and the subsequent removal of alcohol from the albumin are two important elements of albumin production. Table 1 compares alcohol removal from albumin with GH-25 gel at two different flow rates. Increasing the flow rate does not affect de-alcoholization efficiency, albumin recovery or sample dilution. Concentration of the remaining alcohol was below 0.01 % for either flow rate. Cycle time was reduced to one quarter by increasing flow from 29 cm/h/cm˛ to 100 cm/h/cm˛ .

Industrial Desalting:
 
The advantages of desalting biological molecules chromatographically are clearly realized in large-scale applications. From research to pilot and production facility, the scale-up of Cellufine GH-25 has proven to be direct and trouble-free. Mechanical stability of GH-25 allows the use of high flow rates in large columns.

A typical large scale desalting application requiring the processing of 225 liters per day can be accomplished by 5 liters of Cellufine GH-25 in a 105 x 587 mm column (see Table 2). This rigid gel withstands flow rates to 1250 ml/min in that column geometry (870 ml/cm/cm˛). Cycle time between injections is 8 minutes. The high volume load (25 % of Vt), is accommodated by the packing without sacrificing resolution or purity.

 

 

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