IONTOSORB® - Bead Cellulose Derivatives
[ Home ] [ Bead Cellulose with functional groups ] [ Bead Cellulose without functional groups ] [ Index of Publications ] [ Price List ] [ Debtors ] [ MAGNETIC SORBENTS based on Bead Cellulose ]

MT 50
MT 100
MT 200
MT 500
Photos

Macroporous Bead Cellulose - General Information

Macroporous Bead Cellulose MT - Gel Filtration Media for Biomolecule Separations

Macroporous bead cellulose has a hydrophilic matrix and spherical particles. It is produced in several porosity types, marked MT 50,  MT 100,  MT 200  and   MT 500, and various particle size. Macroporous Bead Cellulose, MT is a highly porous regenerated cellulose with heterogenous matrix (partially microcrystalline) and with polymer structure stabilized by hydrogen bonds only - there are no covalent cross-linkings.

One of working sites, which use to deal with a research of bead cellulose applications, was:  Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Science, Bratislava, Slovakia. 

Features

  • Excellent mechanical stability even at gel with large pores. Easy to handle. Resistant to destruction and generation of fine particles during stirring.
  • Rigid spherical particles. High flow throughput the gels at a low drop in pressure.
  • Narrow particle size distribution in several degrees converting all ranges common for this sort of gel.
  • High chemical resistance and compatibility with most commonly used solvents and buffers. Applicability in wide range of pH and salt concentrations with minimal changes (swelling / shrinking) in the gel bed.
  • High temperature stability allows sterilization by autoclaving.
  • High hydrophility. A number of free hydroxy groups in matrix allows preparation of derivatives or enzyme immobilization.
  • High porosity of matrix allows good capacity and recovery.
  • High selectivity of separation.
  • Easy-to-ready. Gels are supplied pre-swollen.

Mechanical Stability

Flow characteristics of  Macroporous Bead Cellulose MT types with various particle sizes

Types Particle Size

[ um ]

 Denotation Flow Performance (1)

[ ml cm-2 hod-1 bar-1 m-1]
MT    50 30 - 50 - 85
50 - 100 Fine 270
100 - 250 Medium > 500
MT   100 30 - 50 - 20
50 - 80 Superfine 170
80 - 100 Fine 300
100 - 250 Medium 600
MT   200 50 - 80 Superfine 130
80 - 100 Fine 220
100 - 250 Medium 450
MT   500 50 - 80 Superfine 45
80 - 100 Fine 80
100 - 250 Medium 250

(1)   For distilled water at 20 oC.

Optimum*) pressure in column during packing with Macroporous Bead Cellulose MT

Types Pressure      [ bar ]
MT    50 > 2.0
MT  100 1.5 - 2.0
MT  200 1.2 - 1.5
MT  500 0.8 - 1.2

*) pressure at which maximum of efficiently is reached

 

Maximum*) flow and pressure throughput for Macroporous Bead Cellulose MT

Types

 Flow  Rate Pressure  [ bar ]
linear   [ cm / hr ] volumetric**)  [ ml/min ]
MT    50 > 760 > 9.9 > 4.0
MT  100 750 9.0 2.2 - 2.7
MT  200 450 6.0 1.5 - 2.0
MT  500 115 1.5 1.0 - 1.5

*) maximum of  flow rate or pressure at which column is stabill not blocked
**) for bead size range 50-80 µm and gel-bed 1 x 60 cm.

 

Total Pore Volume for Various Porozities of Macroporous Bead Cellulose MT

 

Porozity Total Pore Volume Total Pore Volume
[ cm3 / g dry ] [ cm3 / g sucked ]
50 1.3 0.57
100 5.8 0.85
200 8.0 0.89
300 9.2 0.90
500 11.5 0.92

The Pore Volume Determined as W.R.

 

 

Average values of Total Pore Volume for various porozities of Macroporous Bead Cellulose MT

 

 

By their pressure stability Macroporous Bead Cellulose MT belongs to the category of swollen semi-rigid gels for low-pressure chromatography. The gels are characterized by excellent mechanical properties.

 

Flow characteristics of Perloza MT types (3725 bytes)

Flow characteristics of Macroporous Bead Cellulose MT types   Gel bed 1 x 60 cm, particle size 50 - 100 µm for all types

 

Column: 10 x 660 mm,   Eluent: destilled water,   Packing Flow Rate: 760 cm/hr

 

Column: 10 x 660 mm,   Eluent: destilled water,   Packing Pressure: 2.5 bar

 

Column: 10 x 660 mm,   Eluent: destilled water,   Packing Pressure: 1.75 bar

 

Column: 10 x 660 mm,   Eluent: destilled water,   Packing Pressure: 1.25 bar

 

Column: 10 x 660 mm,   Eluent: buffer phosphate 0.2 M   pH = 7,   Packing Pressure: 1 bar

Agarose = agarose gel, Japan = Japan Cellulose 2000

 

Column: 10 x 660 mm,   Eluent: destilled water

Japan = Japan Cellulose 90, 300, 1000,   Dextran = dextran gel 100

 

 

Information on the porosity of Macroporous Bead Cellulose MT materials in water-swollen state as obtained by inverse steric exclusion chromatography (ISEC)

 

Net volume of water-swollen beads*

[ cm3/g ]

Medium pore diameter

[ nm ]
MT    50 1.65 7.4
MT  100 4.86 27
MT  200 6.71 32
MT  300 7.32 38
MT  500 7.43 38

*Does not include the interstitial spaces

The resutls in this table were measured by:

 Ing. Karel Jerabek, Ph.D.

Institute of Chemical Process Fundamentals

Rozvojova 135

165 02  Prague 6

Czech Republic

 

 

Size Exclusion Limits for Properties (Porosity) of Individual MT types

Macroporous Bead Cellulose MT types are highly porous gels well accessible for large macromolecules, as it is shown in this Figures.

 

Selectivity curves of Macroporous Bead Cellulose MT types Kav vs. Molecular Weight of globular proteins in semilogarithmic scale

 

Elution behavior of globular protein on Macroporous Bead Cellulose MT, dependence of distribution coefficient Kav on Molecular Weight

Protein M. W. Kav
[ g / mol ] MT 50 MT 100 MT 200 MT 500
  Blue Dextran 2 000 000 0 0 0.01 0.02
  IgM 1 000 000 0 0.10 0.22 0.36
  Thyroglobulin 660 000 0.01 0.20 0.40 0.50
  Ferritin  *) 440 000 0.01 0.35 0.57 0.65
  Catalase  *) 238 000 0.01 0.47 0.74 0.78
  Aldolase 158 000 0.02 0.55 0.78 0.81
  GLS 135 000 0.05 0.58 0.80 0.82
  Bovine Serum Albumin 66 000 0.19 0.70 0.88 0.87
  Ovalbumin 44 000 0.30 0.75 0.88 0.88
  Chymotrypsinogen A 25 000 0.44 0.81 0.89 0.90
  Myoglobulin 18 000 0.50 0.83 0.90 0.91
  Ribonuclease 13 500 0.55 0.85 0.90 0.92
  Cytochrome C 12 400 0.57 0.86 0.91 0.92
  Bacitracin 1 420 0.71 0.89 0.92 0.95
  L-Histidin 152 0.72 0.90 0.93 0.96
  Alanine 89 0.73 0.91 0.94 0.97

eluent:    buffer 0.2 M phosphate pH 7.0                                 *) buffer 0.013 M phosphate pH 7.0

 

 

 

Selectivity curves of Macroporous Bead Cellulose MT types Kav vs. Molecular Weight of Dextran in semilogarithmic scale

 

Elution behavior of Dextran on Macroporous Bead Cellulose MT, dependence of distribution coefficient Kav on Molecular Weight

DEXTRAN

Molecular Weight

 Kav
MT 50 MT 100 MT 200 MT 300 MT 500
1 500 0.45 0.94 0.93 0.99 0.99
6 000 0.18 0.79 0.81 0.98 0.98
10 000 0.01 0.72 0.75 0.84 0.83
17 500 0 0.62 0.67 0.76 0.76
40 000 0 0.45 0.52 0.63 0.62
70 000 0 0.31 0.39 0.51 0.50
110 000 0 0.20 0.28 0.41 0.39
196 300 0 0.07 0.15 0.26 0.25
400 000 0 0 0.03 0.09 0.08
2 000 000 0 0 0 0 0

 

 

It is necessary to mention that the pore volume accessible to a given molecule is determined by its size (weight) and the pore size distribution of packing and can be defined by the following formula:

vzorec.gif (1272 bytes)

where Kav is the distribution coefficient (the factor of pore volume available for a given molecule), Ve is elution (retention) volume, Vo is void (interstitial) volume and Vt is total column volume.

 

Selectivity parameters of Macroporous Bead Cellulose MT

Types Exclusion Limit Fractionation Range
MW proteins,  g / mol
MT  50 1 x 105 20 000   -   70 000
MT  100 2 x 106 30 000   -   1 000 000
MT  200 4 x 106 200 000   -   2 000 000
MT  500 > 5 x 106 300 000   -   4 000 000

 

Types Bead Volume [ ml /g dry ] Exclusion Limit  [MW dextrans ]
MT  50 3 - 5 50 000
MT  100 9 - 11 100 000
MT  200 13 - 16 200 000
MT  500 18 - 21 500 000

 

Other Technical Data

Temperature Resistance (2)  oC :     120

Stability within pH Range :               1 - 14            NaOH  or   HCl  (c = 1 mol / l)

Stability in Salt Solutions :                with ionic strength up to 10 mol/l

Swelling / Shrinkage (vol %) :
      with changes in pH, salt concentration (3)                                               1 max.
      with replacing of aqueous solutions by polar organic solvents                 10 max.

Chemical Resistance :       aqueous solutions, buffer, organics, detergents
                                            and chaotropic agents (including SDS, 8 M urea)

(2)    In wet state, pH 7.0  for 1 hour.
(3)    Over the whole range of pH anionic strength.

 

Handling with Macroporous Bead Cellulose and Sorbents IONTOSORB

Bead Cellulose and its derivatives must not be dried, because drying results in loss of porosity. Therefore they are supplied fully swollen in water, sucked off. They can be stabilized by sodium azide (0.02 %) or ethanol (20 %) according to customer´s demand.