GE AKTApure, User Manual, Page: 442 / 522

Included phases Principle Predefined purification
method
After equilibration and sample application, the
proteins are eluted isocratically. This technique
is commonly used for buffer exchange. Desalting
After equilibration and sample application, pro-
teins separate and elute according to their size
(largest first). Gel filtration (GF)
After equilibration and sample application (use
a buffer containing a high salt concentration,
for example 2 M ammonium sulfate) hydropho-
bic proteins are adsorbed to the column ligand.
After a wash to remove unbound sample, elution
is performed using a gradient of decreasing salt
concentration. Finally, the column is washed and
re-equilibrated with start buffer. Hydrophobic Interaction
Chromatography (HIC)
The sample application loops of the loop valve
are manually filled with samples. The method
will guide the user through the process by
pausing and displaying on-screen instructions.
Up to five loops can be filled with different sam-
ples. The loops are filled in descending order.
Partial or complete loop fill can be chosen. Auto-
matic washing of loops and flow path are inte-
grated in the method. Manual Loop Fill
The required components for this phase are a
column valve and a loop valve.
442 ÄKTA pure User Manual 29119969 AB
9 Reference information
9.7 Predefined methods and phases
9.7.1 Predefined purification methods