GE ÄKTA start Manual page 9

The pre-set run parameters for each technique are listed in the table below.
Parameter AC step
HiTrap
Column Volume
(ml)
Flow rate
(ml/min)
Pressure Limit
(MPa)
Save Result to
USB *
Sample From
Sample Volume
(ml) **
Equilibration
Volume (CV)
Wash Unbound
Volume (CV)
Elution Option Isocratic
Target B Conc
(%)
Elution Volume
(CV)
Hold at 100% B
(CV)
Re-Equilibration
Volume (CV)
Fractionation
Volume (ml)
* 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.
ÄKTA start System Cue Card, 29-0240-42 AC
AC step DS 5 ml
1 ml 5 ml HiTrap
HiTrap
1.0 5.0
1.0 5.0
0.3 0.3
Yes Yes
Pump Pump Pump
0.1** 0.1** 0.1**
5.0 5.0
15.0 15.0
Isocratic Isocratic
100 100
5.0 5.0
NA NA
5.0 5.0
1.0 3.0
DS IEX step
53 ml 1 ml
HiPrep™ HiTrap
5.0 53.0 1.0
3.0 5.0 1.0
0.3 0.2 0.3
Yes Yes Yes
Pump Pump
0.1** 0.1**
3.0 3.0 5.0
NA NA 10.0
Isocratic Isocratic
NA NA 100
1.2 1.2 5.0
NA NA NA
NA NA 5.0
0.5 3.0 1.0
%B
100
50
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.
IEX step IEX
5 ml gradient
HiTrap 1 ml
HiTrap
5.0 1.0
5.0 1.0
0.3 0.3
Yes Yes
Pump Pump
0.1** 0.1**
5.0 5.0
10.0 10.0
Isocratic Gradient
100 100
5.0 20.0
NA 5.0
5.0 5.0
3.0 1.0
5 CV 15 CV
IEX GF
gradient 16/60
5 ml HiPrep
HiTrap
5.0 120
5.0 1.0
0.3 0.2
Yes Yes
Pump Pump
0.1** 0.1**
5.0 0.2
10.0 NA
Gradient Isocratic
100 NA
20.0 1.2
5.0 NA
5.0 NA
5.0 4.0
Fractionation/
collection
5 CV 5 CV
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