ÄKTA start System Cue Card, 29-0240-42 AC
9
The pre-set run parameters for each technique are listed in the table below.
Parameter
AC step
1 ml
HiTrap
AC step
5 ml
HiTrap
DS 5 ml
HiTrap
DS
53 ml
HiPrep™
IEX step
1 ml
HiTrap
IEX step
5 ml
HiTrap
IEX
gradient
1 ml
HiTrap
IEX
gradient
5 ml
HiTrap
GF
16/60
HiPrep
Column Volume
(ml)
1.0
5.0
5.0
53.0
1.0
5.0
1.0
5.0
120
Flow rate
(ml/min)
1.0
5.0
3.0
5.0
1.0
5.0
1.0
5.0
1.0
Pressure Limit
(MPa)
0.3
0.3
0.3
0.2
0.3
0.3
0.3
0.3
0.2
Save Result to
USB *
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Sample From
Pump
Pump
Pump
Pump
Pump
Pump
Pump
Pump
Pump
Sample Volume
(ml) **
0.1**
0.1**
0.1**
0.1**
0.1**
0.1**
0.1**
0.1**
0.1**
Equilibration
Volume (CV)
5.0
5.0
3.0
3.0
5.0
5.0
5.0
5.0
0.2
Wash Unbound
Volume (CV)
15.0
15.0
NA
NA
10.0
10.0
10.0
10.0
NA
Elution Option
Isocratic Isocratic Isocratic Isocratic Isocratic Isocratic Gradient Gradient Isocratic
Target B Conc
(%)
100
100
NA
NA
100
100
100
100
NA
Elution Volume
(CV)
5.0
5.0
1.2
1.2
5.0
5.0
20.0
20.0
1.2
Hold at 100% B
(CV)
NA
NA
NA
NA
NA
NA
5.0
5.0
NA
Re-Equilibration
Volume (CV)
5.0
5.0
NA
NA
5.0
5.0
5.0
5.0
NA
Fractionation
Volume (ml)
1.0
3.0
0.5
3.0
1.0
3.0
1.0
5.0
4.0
* the results will be saved only if the USB memory stick is
inserted before opening the method.
** variable parameter
Note: The sample can be loaded only through Pump.
Quick start methods description
The “ready to run” methods available with ÄKTA start
are presented below. The length of each step or phase is
indicated on the diagrams as column volumes (CV).
Sample:
variable volume to be set by the user.
Pump wash:
- for AC/IEX: 5 ml Buffer B, then 5 ml of Buffer A wash.
- for DS/GF: 5ml of Buffer A wash.
AC Step 1 ml/5 ml HiTrap - Affinity Chromatography
Commonly used for purification of tagged proteins, e.g.,
His tag, GST tag, Mab tagged proteins, etc.
The bound proteins are eluted in a single step using
single elution buffer.
•
•
Re
-e
qu
ili
br
at
io
n
W
as
h
un
bo
un
d
El
ut
io
n
Pu
m
p
w
as
h
Eq
ui
lib
ra
tio
n
100
50
5 CV
15 CV
5 CV
5 CV
Sa
m
pl
e
%B
Fractionation/
collection
DS 5 ml HiTrap/53 ml HiPrep - Desalting/Buffer Exchange
Chromatography
The proteins are eluted in a single step using single
elution buffer.
Make sure to load the recommended sample volume.
•
•
