114
BULK
TOYOPEARL AND TSKgel BULK RESINS FOR HIC
HIGHLIGHTS
A wide range of hydrophobicities is suitable for most proteins.
Standard 100 nm pore size is available for large biopolymers, and
three Butyl pore sizes (50 nm, 75 nm and 100 nm) are available.
TOYOPEARL “600M” series of HIC resins with optimized pore size
of 75 nm for antibody separation. Phenyl-600M and Butyl-600M with
highest DBCs for IgG.
Seamless scale up from high efficiency TSKgel 5PW-type analytical
columns is possible.
Hydrophobic Interaction Chromatography (HIC) has become
a popular mode of chromatography for the purification of biopolymers
at analytical as well as preparative scale. This is accomplished by
the interaction of hydrophobic ligands on the base matrix with the
hydrophobic areas located on the surface of proteins. HIC is an excellent
complement to size exclusion and ion exchange chromatography in
difficult separations, particularly those where the contaminants are of
similar pI or molecular weight. It is often preferred over reversed phase
chromatography when preservation of biological activity of the protein
is of utmost importance.
Tosoh Bioscience offers both the TSKgel and TOYOPEARL resin product
lines for HIC. See
Table IV
for a complete listing of functionalities.
Each product line has similar backbone chemistry. TSKgel 5PW-type
resins possess a higher degree of cross-linking than the corresponding
TOYOPEARL resins. Additionally, choices in particle size are offered to
match the desired resolution and throughput. A variety of HIC bulk media
are offered as LABPAK kits in quantities < 1 L and in a combination of
resins with varying functionalities. Additionally, HIC media are available
in ToyoScreen process development columns for convenient scouting
and methods development.
Ordering information for quantities < 1 L is provided at the end of this
section.
Description
Strength
*
Part. size
grades (µm)
TSKgel Ether-5PW
1
20, 30
TOYOPEARL Ether-650
1
35, 65
TOYOPEARL PPG-600
2
65, 100
TSKgel Phenyl-5PW
3
20, 30
TOYOPEARL Phenyl-650
3
35, 65, 100
TOYOPEARL Phenyl-600
4
65
TOYOPEARL Butyl-600
4
65
TOYOPEARL Butyl-650
4
35, 65, 100
TOYOPEARL SuperButyl-550
4
100
TOYOPEARL Hexyl-650
5
100
* Relative scale: 1 = least hydrophobic, 5 = most hydrophobic.
Applications:
HIC resins can be applied to separate/purifiy proteins with similar
chemical or structural properties, plasmids and monoclonal antibodies.
See
FIGURE 9
for separation of large glycoprotein from crude extract on
TOYOPEARLButyl-650S. Please visit ourwebsite:
.
de for extensive application data.
Column: TOYOPEARL Butyl-650S, 22 mm ID x 26 cm L;
Sample: crude protein from sea hare Aplysia kurodai;
Elution: multi-step (NH
4
)
2
SO
4
in 50 mmol/L phosphate buffer,
pH 7.0 A. load & wash: 40 % saturated (NH
4
)
2
SO
4
B. 20% saturated (NH
4
)
2
SO
4
C. 0% saturated (NH
4
)
2
SO
4
figure 9
Large glycoprotein purified on TOYOPEARL Butyl-650S
Large glycoprotein purified on Toyopearl Butyl-650S
Fraction Number (8mL/fraction)
Absorbance, 280nm
Antibacterial activity (%-Inhibition)
2.0
1.0
0
20
10
30
70
100
Column:
Sample:
El tio :
Toyopearl Butyl-650S, 22mm ID x 26cm
crude protein from sea hare Aplysia kurodai
multi-step (NH
4
)
2
SO
4
in 50mmol/L phosphate buffer, pH 7.0
A. load & wash: 40% s turat d (NH
4
)
2
SO
4
B. 20% saturated (NH
4
)
2
SO
4
C. 0% saturated (NH
4
)
2
SO
4
40
50 60
80 90
A.
C.
B.
100
50
0
TABLe IV
TOYOPEARL and TSKgel HIC resins
