Bio-Rad ChromLab™ Software - Manual (Page 291)

Multicolumn Purification Method Templates
User Guide | 289
Multicolumn Tandem Purification Templates
Tandem purifications are used when two different chromatography techniques are
combined into one method. First, samples are injected, bound to a column and
washed to remove any major contaminants. Next, the target fractions are eluted and
either applied directly onto another column in tandem or stored temporarily in a
sample loop or container. If stored, they are later reinjected onto a second column.
In either case, the second column is then eluted and the purified fractions are
collected with the BioFrac fraction collector or an outlet valve. ChromLab software’s
multicolumn tandem purification templates utilize affinity, desalting, and size
exclusion chromatography (SEC) techniques but can be modified to include other
techniques.
Table 8 defines the multicolumn tandem purification templates.
Table 8. Multicolumn tandem templates
Template Name Description
2-D Affinity (1 ml) > SEC (24 ml) This method is used to purify up to four samples. Using a
sample inlet valve and the sample pump, the method
loads different samples onto separate affinity columns
and washes unbound material off in quick succession.
This process may be repeated for up to four columns. This
method is designed to reduce the target protein’s
exposure to proteases. The target proteins are then eluted
from each column sequentially using a step gradient. The
eluted fraction is stored in a loop valve. The fraction is
then injected onto a size exclusion column for a final
analytical or preparative purification step.