sec
SEC
13
TSKgel SW-type Gel Filtration columns
Proteins (general)
Choose one of the TSKgel SW
XL
columns using the calibration curves
on
page 12
to select the appropriate pore size based on knowledge or
estimate of protein size.
Monoclonal antibodies
TSKgel SuperSW mAb columns have been developed for the analysis
of monoclonal antibodies. They provide higher resolution (HR) or faster
analysis (HTP) than the TSKgel G3000SW
XL
which is traditionally used
for quality control in many QC labs. TSKgel SuperSW3000 is utilized
when sample is limited or at very low concentration.
Peptides
TSKgel G2000SW
XL
is the first selection for the analysis of peptides.
TSKgel SuperSW2000 is utilized when sample is limited or at very low
concentration.
Other
The use of TSKgel SuperSW columns requires optimization of the HPLC
system with respect to extra-column band broadening. Capillary tubing
ID, injection volume, detector cell volume, and detector time constant
all need to be reduced to fully benefit from the high column efficiency
and small peak volumes of the SuperSW columns. Use SW columns
when not sample limited or when larger amounts of sample need to be
isolated.
Comparing TSKgel SW, SW
XL
and SuperSW Gel Filtration
Columns
Figure 1
&
Figure 2
show the increased resolution and sensitivity of
the TSKgel SuperSW columns compared to TSKgel SW
XL
columns. This
is due to the smaller particle size (4 vs. 5 µm) and the narrow column
diameter (4.6 mm ID).
Comparison of TSKgel SuperSW3000 and TSKgel G3000SW
XL
for the separation of proteins
Minutes
Column:
Sample:
Elution:
Flow Rate:
Temp:
Detection:
A.TSKgel G3000SW
XL
, 7.8mm ID x 30cm;
B. TSKgel SuperSW3000, 4.6mm ID x 30cm
5µL of a mixture of 1. thyroglobulin, 0.5mg/mL (660,000Da);
5. p-aminobenzoic acid, 0.01mg/mL (137Da)
0.1mol/L NaSO
4
in 0.1mol/L in phosphate buffer with 0.05%
NaN
3
, pH 6.7
1.0mL/min for G3000SW
XL
; 0.35mL/min for SuperSW3000
25
°
C
UV @ 220nm
0
10
20
30
40
50
3 5 7 9 11 13 15
0
10
20
30
40
50
5
Rs: 10.7
4
Rs: 4.0
2
3
Rs: 3.5
1
5
4
3 2
1
RS: 8.4
RS: 3.0
RS: 3.3
A. G3000SW
XL
B. SuperSW3000
mV
4. ribonuclease A, 1.5mg/mL (12,600Da);
Minutes
3 5 7 9 11 13 15
2.
g
-globulin, 1.0mg/mL; (150,000 Da);
3. ovalbumin, 1.0mg/mL (43,000Da);
Column: A. TSKgel G3000SW
XL
, 7.8 mm ID x 30 cm L;
B. TSKgel SuperSW3000, 4.6 mm ID x 30 cm L;
Sample: 5
μ
L of a mixture of 1. thyroglobulin, 0.5 mg/mL (660,000 Da);
2.
g
-globulin, 1.0 mg/mL (150, 00 Da); 3. ovalbumin, 1.0 mg/mL (43,000 Da);
4. ribonuclease A, 1.5 mg/mL (12,600 Da); 5.
r
-aminobenzoic acid,
0.01 mg/mL (137 Da);
Elution: 0.1 mol/L NaSO
4
in 0.1 mol/L in phosphate buffer with 0.05 % NaN
3
,
pH 6.7; Flow rate: 1.0 mL/min for G3000SW
XL
; 0.35 mL/min for SuperSW3000;
Temp: 25°C; Detection: UV @ 220 nm
figure 1
Comparison of tskgel super sw3000 and tskgel g3000sw
XL
for the
separation f proteins
3
5
7
9 11 13 15
0
10
20
30
40
50
60
Minutes
mv
column:
Sample:
Inj. volume:
Elution:
Flow rate:
temp:
detection:
A. tSKgel G2000SW
XL
, 7.8mm Id x 30cm;
B. tSKgel SuperSW2000, 4.6mm Id x 30cm
1. thyroglobulin (0.2mg/mL); 2. albumin (1.0mg/mL);
3. ribonuclease A (1.0mg/mL); 4. p-aminobenzoic acid (0.01mg/mL)
5µL
0.1mol/L phosphate buffer + 0.1mol/L na
2
So
4
+ 0.05% nan
3
(pH 6.7)
0.35mL/min for SuperSW2000; 1.0mL/min for G2000SW
XL
25
°
c
uv @ 280nm
1
rs:4.7 rs:4.6
rs:11.9
2
3
4
G2000SW
XL
3
5
7
9 11 13 15
0
10
20
30
40
50
60
Minutes
mv
1
rs:4.8
rs:6.2
rs:15.6
2
3
4
SuperSW2000
Comparison of TSKgel SuperSW2000 and TSKgel G2000SW
XL
for the separation of proteins
A.
B.
Column: A. TSKgel G2000SW
XL
, 7.8 mm ID x 30 cm L;
B. TSKgel SuperSW2000, 4.6 m ID x 30cm L;
Sample: 1. thy oglobulin (0.2 mg/ L); 2. albumin (1.0 mg/mL); 3. ribonuclea-
se A (1.0 mg/mL); 4. p-aminobenzoic acid (0.01 mg/mL);
Inj. Volume: 5 μL; Elution: 0.1 mol/L phosphate buffer + 0.1 mol/L Na
2
SO
4
+
0.05 % NaN
3
(pH 6.7);
Flow rate: 0.35 mL/min for SuperSW2000; 1.0 mL/min for G2000SW
XL
;
Temp: 25°C; Detection: UV @ 280 nm
figure 2
Comparison of tskgel super sw2000 and tskgel g3000sw for the se-
paration of Proteins
