Bio-Rad ChromLab™ Software - Manual (Page 164)

6 | Creating a Method
162 | NGC Chromatography Systems and ChromLab Software
Multicolumn Tandem Multicolumn tandem purification uses these templates
when two different chromatography techniques are
combined into one method. First, samples are injected,
bound to a column and washed to remove any main
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 then re-injected 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.
Reverse Phase Reverse phase chromatography utilizes hydrophobic regions
on target proteins to bind to a hydrophobic column matrix. A
gradient of increasing organic solvent concentration/ratio is
used to elute the proteins from the column matrix.
Size Exclusion (Gel Filtration) Size exclusion/gel filtration chromatography separates
proteins based on their molecular size or weight. Proteins do
not bind to the column matrix and are typically eluted
isocratically. Select a buffer system that maximizes the
stability of the target protein.
System Test These methods test the performance of NGC systems that
have a UV detector. They are intended to assess the flow rate
and gradient accuracy of the system pumps, the functionality
of the mixer, and the responses of the UV and conductivity
detectors. In most cases, a visual examination of the resulting
chromatogram can confirm proper performance.
Table 6. Standard Method Templates, continued
Method Template Explanation