Iec
IEC
53
TSKgel CATION EXCHANGE COLUMNS
HIGHLIGHTS
TSKgel SP-STAT and CM-STAT nonporous columns provide high
efficiency separation at short analysis time.
Pore structure and bonding chemistry of TSKgel BioAssist S
provides high capacity for medium to large MW proteins.
BioAssist columns are packed in 4.6 mm ID or 10 mm ID PEEK
hardware. Other columns are available in glass and stainless steel
for analytical, semi-preparative and preparative applications.
Binding capacity for small to medium size proteins on TSKgel
CM-3SW is approximately double that of TSKgel CM-5PW due to the
smaller pore size and larger surface area.
The TSKgel SP-5PW column is available in 2 mm ID format for
LC-MS applications.
The basic properties of TSKgel STAT cation exchange columns are
summarized in Table II
APPLICATIONS
TSKgel SP-STAT, CM-STAT
Nonporous TSKgel STAT columns provide fast, high resolution
separations at moderate pressures.
Figure 9
shows the monitoring of
a PEGylation reaction of beta-lactoglobulin on a short SP-STAT column.
TSKgel CM-STAT
columns are ideally suited to analyze the profile of
charge isomers of proteins.
FIGURE 10
shows the analysis profiles for
five antibodies and their charge isomers separated on a TSKgel CM-
STAT column.
0
20
10
30
40
Retention time (min)
40
30
20
10
0
mV
A
B
C
D
E
Column: TSKgel CM-STAT column (7 μm, 4.6 mm ID x 10 cm L); Flow rate:
1 mL/min; Mobile phase: A: 20 mM MES (pH 6.0), B: 20 mM MES + 0.5 M
NaCl (pH 6.0); Gradient 10% B to 15 % B in 15 minutes; Detection: UV @ 280
nm, Injection volume 20 μL
figure 10
Separation of MAB charge variants on TSKgel CM-STAT
Column: Prototype SP-STAT, 4.6 mm ID x 3.5 cm L, (10 um)
Eluent: A: 20 mmol/L Na acetate buffer pH 4.5, B: 0.8 mol/L NaCl in A pH 4.5;
Gradient: 0 to 30% B (2 min); Flow rate: 4.0 mL/min; Detection: UV @ 280 nm
Real-time analysis of PEGylation reaction (PEG MW=5000) at 5-minutes
intervals
figure 9
Monotoring of PEGylation of -lactoglobulin
Property
TSKgel SP-STAT
TSKgel CM-STAT
Base
material
Cross-linked hydrophilic polymer (mono-disperse
particles)
Pore size
Non-porous
Functional
group
Sulfonate
Carboxymethyl
Particle
size
7 µm
10 µm 7 µm 10 µm
Column
size
4.6 mm ID x
10 cm L
3 mm ID x
3.5 cm L
4.6 mm ID x
10 cm L
3 mm ID x
3.5 cm L
Applica-
tion
High resolu-
tion protein
separation
High throughput protein separation
TABLE I I
Basic Properties of TSKgel STAT cation exchange Columns
Minutes
