Bio-Rad Nuvia™ cPrime™ Media - Manual (Page 7)

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5. Using a compression factor of 1.17, calculate the volume of
slurry required for the intended bed height.
a. For example, for a 20 cm bed height using a 50% slurry, the
volume would be:
1.17 (20/0.5) * πr
2
6. Add a small amount of packing buffer to the column to wet the
bottom frit, then pour in calculated amount of resin slurry.
7. Insert the column flow adaptor and flow pack at a linear velocity of
300–600 cm/hr with packing buffer for at least 10 min. Note the
compressed bed height, stop the flow, and adjust the flow adaptor to
compress the bed to the intended bed height.
8. Equilibrate with at least 3 column volumes (CV) of equilibration
buffer and evaluate column efficiency using your standard operating
procedures or the procedure described in Section 5.
Packing Process Scale Columns
After removing the storage buffer (Section 3), prepare a 20–50%
slurry (v/v) with packing buffer (see Table 2). For most process
columns, follow the manufacturer’s recommendations with one
major exception: do not recirculate the Nuvia cPrime media slurry
through the packing pump. Use a low-shear impeller for automatic
mixing or a plastic paddle for manual mixing to avoid damaging the
media. The best overall performance of Nuvia cPrime media will
be obtained with a compression ratio of 1.15–1.20. Compression
factor is defined as settled bed height divided by packed bed
height.
After achieving the desired compression ratio, it is recommended
to condition the column by flowing fresh packing or equilibration
buffer for 3 CV followed by 3 CV in downflow at the chosen
process flow rate. After this flow conditioning step, evaluate
column efficiency using your standard operating procedures or the
procedure described in Section 5.