Bio-Rad Media Sampler Pack - Manual (Page 8)

column is reached, or to the flow rate at which the maximum pressure for
Macro-Prep is reached (7 bar or 102 psi). As the bed compresses with
increasing flow rate, stop the flow and adjust the flow adaptor so it penetrates
1–2 mm into the media bed. Repeat the flow and adjustment cycle until the
bed no longer compresses.
Note: Always pack the column at the highest flow rate your chromatography
system permits, not exceeding the recommended maximum linear flow rate of
3,000 cm/hr. A well-packed bed will provide better resolution.
7. Attach the column to your chromatography system and purge with the
starting buffer at the maximum flow rate to be used during operation. You
should not see any further bed compression. If you do, adjust the flow
adaptor 1–2 mm into the media bed.
8. Equilibrate the column in starting buffer. The column is now ready for
sample application.
Note: For optimal long-term use, do not exceed 80% of the maximum pressure
attained during the column packing.
Packing Large Columns
In large columns, Macro-Prep media should be packed using a 50% slurry at constant
pressure up to a maximum of 7 bar (102 psi). Pack with a constant flow rate that is
approximately 25% greater than the anticipated maximum to be used during operation.
Given the variety of industrial column hardware and packing skids, we recommend using
standard operating procedures for packing provided by the column vendor. Bio-Rad
EasyPack
™ and GelTec ™ columns are supplied with a movable piston. The GelTec ™
column is packed by pumping the packing solution through the media bed at constant
pressure or flow.
1. Packing begins with preparation of the tubing and the column. Clean and rinse the
column well with packing buffer before packing. The rinse is also used to displace
air from the column and its exit tubing. Make sure there is no air trapped in the
bottom flow cell. After the air has been removed from the bottom flow cell and exit
tubing, leave about 2 cm of liquid in the bottom of the column. Close the outlet
valve(s).
2. In a suitable container, mix the packing buffer with the media to form a 50% slurry.
3. Pour or pump the slurry into the column.
4. Insert the movable piston and lower it to the surface of the liquid, making sure no air
is trapped under the piston.
5. Pressurize the inflatable seal to 2 bar (29 psi), so that as the piston is lowered into
the slurry, the buffer exits the top of the column through the inlet line. Be sure the
inlet line is completely filled with packing buffer to avoid air being pumped back into
the column.
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