Bio-Rad Media Sampler Pack - Manual (Page 6)

Packing Small Columns
The following slurry packing method was designed to pack 25 ml of Macro-Prep
HIC media into 5–15 mm ID, low- to medium-pressure lab columns. All buffers
should be degassed prior to packing. Use a packing reservoir that holds ~100 ml.
1. Prepare degassed buffer or water to be used as the packing solution.
2. Macro-Prep is shipped as a 50% slurry, so measure 50 ml of suspended
slurry into a 100 ml graduated cylinder. Allow the media to settle. Decant the
shipping solution from the settled media.
3. Add 25–50 ml of degassed packing buffer to the media.
4. Seal the cylinder and rotate it to suspend the media. Caution: Do not mix with
a magnetic stir bar, to avoid formation of small fines. Larger volumes of slurry
may be mixed with an overhead stirrer at low to moderate speed.
5. Fill the column with approximately 2 cm of packing buffer. Pour all of the
media slurry into the column with the packing reservoir attached.
6. Insert the column flow adaptor and start flow at ~100 cm/hr. Gradually
increase the flow rate from 100 cm/hr to the flow rate at which the maximum
allowable pressure for the 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.
7. Attach the column to your chromatography system and purge with 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. The column is now ready for use.
Note: For optimal long-term use, do not exceed 75% of the maximum
pressure attained during the column packing.
Packing Large Columns
In large columns, Macro-Prep 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 use of standard operating procedures for packing provided by the
column vendor.
Bio-Rad EasyPack ™ and GelTec ™ columns are supplied with a movable piston.
They are packed by pumping the packing solution through the media bed at
constant pressure or flow.
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