23
IEC
Impact of pore size and ligand attachment on
DBC
Table III contains DBC data for five of our TOYOPEARL
resins using three different sized proteins. There are three
different pore sizes and three different ligand attachment
methods represented. TOYOPEARL GigaCap Q-650M
has the highest capacity for all combinations of pore size
and attachment chemistries. Please note the decrease in
capacity for the larger proteins on the TOYOPEARL SuperQ-
650M resin indicating that the accessible pore volume has
diminished by the ligand attachment chemistry used.
Resin Pressure Flow Properties
All TOYOPEARL resins are designed to withstand pressures
up to 3 bar. TOYOPEARL GigaCap resins have a particle size
of 50-100 microns which is slightly larger than our normal
M-grade 40-90 micron beads. This particle size difference
generates a lower back pressure (Figure 8) than our more
traditional M-grade ion exchange products. The TSKgel
5PW type resins can be operated at pressures up to
20 bar.
If recommended packing procedures are followed,
TOYOPEARL and TSKgel IEC resins maintain stable bed
volumes during the pH and ionic strength changes that
occur during normal ion exchange chromatography
(Consult our TOYOPEARL Instruction Manual for the recom-
mended packing conditions).
Multi-cycle gradient operation and re-equilibration are
accomplished without volume changes in the TOYOPEARL
column bed. The mechanical stability of the resins allows
the use of longer column beds with more efficiency or
higher operational flow rates.
Ion exchange
chromatography
figure 8
0
1
2
3
0
200
400
600
800
1000 1200 1400
Pressure (Bar)
Linear Velocity (cm/h)
TOYOPEARL GigaCap S-650M
TOYOPEARL GigaCap Q-650M
TOYOPEARL GigaCap CM-650M
TOYOPEARL SuperQ-650M
TOYOPEARL SP650M
Pressure-flow curve comparison
Column size:
22 mm ID x 20 cm
Mobile phase:
distilled water
Temperature:
25 °C
Column size: 22 mm ID x 20 cm L; Mobile phase: distilled water
Temperature: 25 °C
Pressure-flow curve comparison
TABLE III
Column size: 6 mm ID x 4 cm L; Sample concentration: 1 mg/mL; Loading buffers: BSA 0.05 mol/L Tris-HCl (pH = 8.5); Human IgG 0.05 mol/L Tris-HCl (pH
= 8.7); Thyroglobulin 0.05 mol/L Tris-HCl (pH = 8.7) + 0.15 mol/L NaCl
Elution buffers: loading buffer + 1.0 mol/L NaCl; Flow rate: 212 cm/h; Detection: UV @ 280 nm
Dynamic binding capacity varies with protein size
BINDING CAPACITY (g/L gel)
RESIN
PORE
BSA
HUMAN IgG THYROGLOBULIN
SIZE (nm)
66 kDa
160 kDa
660 kDa
TOYOPEARL GigaCap Q-650M
100
173
108
71
TOYOPEARL SuperQ-650M
100
145
13
3
TOYOPEARL Q-600C AR
75
108
90
26
TOYOPEARL QAE-550C
50
29
32
6
TOYOPEARL DEAE-650M
100
25
31
3
