8
IEC
IEC
TSKgel STAT SERIES
Applications with TSKgel STAT Anion Exchange
Columns
Monoclonal Antibodies
The monoclonal antibody was digested using pepsin and
separated on a TSKgel Q-STAT column and a competi-
tive non-porous WAX column. As shown in Figure 5, three
peaks were isolated from the TSKgel Q-STAT column and
assigned as F(ab‘)2, pFc and intact IgG by SDS-PAGE. There
was no correlation between the peaks obtained on the com-
petitive WAX column and SDS-PAGE.
High Resolution versus high Throughput Analysis of
Nucleotides
The separation of nucleotides on a 10 cm length Q-STAT
column was compared to the separation on a 3.5 cm length
column. While the longer column provides a high resolution
separation of the nucleotides within five minutes the same
sample can be analyzed within one minute on the shorter
column allowing high throughput. The chromatograms are
shown in Figure 6.
0
2
4
6
8
10
mV
-
reduced SDS
200kDa
116kDa
14.4kDa
6.5kDa
(A)
(B)
Retention time (min)
Non
-PAGE
Intact IgG
F(ab’)
2
pFc
Columns:
A: TSKgel Q-STAT, 7µm, 4.6mm ID x 10cm
B: ProPac WAX-10, 10µm, 4mm ID x 25cm
Eluent:
A: 20mmol/L Tris-HCl (pH8.5)
B: 0.5mol/L NaCl in buffer A
Gradient:
0% B (0min), 100% B (10min)
Flow-rate:
1.0mL/min
Detection:
UV@280nm
Sample:
pepsin digested mAb
figure 5
Column: A: TSKgel Q-STAT, 7 µm, 4.6 mm ID x cm L: B: Competitor WAX,
10 µm, 4 mm ID x 25 cm L; Eluent: A: 20 mmol/l Tris-HCI (pH 8.5); B: 0.5
mol/L NaCl in buffer A; Gradient: 0% B (0 min), 100% B (10 min); Flow rate:
1.0 mL/min; Detection: UV @ 280 nm; Samples: pepsin digested mAb
High resolution:
Column: Q-STAT, 4.6 mm ID x 10 cm L (7 µm); Eluent: A) 20 mmol/L
Tris-HCl (pH 8.5) B) 0.5 mol/L NaCl in A (pH 8.5); Gradient: 0 to 100% B
(10 min.); Flow rate: 1.5 mL/min.; Detection: UV @ 260 nm
High throughput:
Column: Q-STAT, 4.6 mm ID x 3.5 cm L (10 µm); Eluent: A) 20 mmol/L
Tris-HCl (pH 8.5), B) 0.5 mol/L NaCl in A (pH 8.5); Gradient: 0 to 100% B
(1 min.); Flow rate: 4.0 mL/min.; Detection: UV @ 260 nm
Figure 6
high resolution versus high throughput analysis of nucleotides
Analysis of IgG fragments
