GE AKTApure, User Manual, Page: 440 / 522

Included phases Principle Predefined purification
method
After equilibration, the sample of tagged protein
is applied. After washing to remove unbound
sample the column is equilibrated with cleavage
buffer, a protease is applied and the flow is
stopped. The cleaved protein and the protease
are eluted by starting the flow. The next step
regenerates the column, eluting the tag, un-
cleaved protein and protease (if tagged). Finally,
the column is re-equilibrated.
Required components
The required components for this method are
Inlet valve (2-ports or 7-ports) and Mixer valve,
or the sample pump.
Required solutions
The method phases are pre-configured to use
the following solutions in the following inlets and
loop:
Inlet A1: Equilibration buffer
Inlet A2 or sample inlet: Sample
Inlet B1: Cleavage buffer
Inlet B2: Affinity Regeneration buffer
Loop: Protease
Affinity Chromatography (AC)
with Tag Removal
Note:
• Select Inject sample directly onto the col-
umn in the sample application phase.
• The protease application phase is preconfig-
ured to empty the loop with the protease with
a volume of 0.7 ml. The recommended pro-
tease volume is 0.7 CV (column volume). If a
column volume different than 1 ml is used,
this value needs to be changed to correspond
to 0.7 CV.
440 ÄKTA pure User Manual 29119969 AB
9 Reference information
9.7 Predefined methods and phases
9.7.1 Predefined purification methods