GE AKTA avant User Manual page 452

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Table of Contents

9 Reference information

9.6 Predefined methods and phases

Predefined purification

Hydrophobic Interaction

Chromatography (HIC)

Manual Loop Fill

After equilibration and sample application

(use a buffer containing a high salt concen-

tration, e.g, 2 M Ammonium Sulphate) hy-

drophobic proteins are adsorbed to the

column ligand. After a wash to remove

unbound sample, elution is performed us-

ing a gradient of decreasing salt concen-

tration. Then, the column is washed and

re-equilibrated with start buffer. After col-

umn re-equilibration, a fraction collector

accumulator wash is done to minimize the

risk for salt precipitation.

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 samples. The loops

are filled in descending order. Partial or

complete loop fill can be chosen. Automat-

ic washing of loops and flow path are inte-

grated in the method.

The required components for this phase

are a column valve and a loop valve.

Included phases

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