Bio-Rad Nuvia™ Q Media - Manual (Page 6)

Section 3 : Preparation
Nuvia ™ media are supplied fully hydrated in 20% ethanol + 1 M NaCI as
a 50% (v/v) slurry. For column packing, removal of the shipping buffer
is recommended. Small volumes of Nuvia media are easily washed in a
Büchner funnel with 4–5 volumes of packing buffer. For large volumes,
cycling through 3–4 settling and decanting steps using the packing buffer
in the shipping container is recommended.
Removal of fines from Nuvia media is not required due to the narrow
particle size range. If fines have been generated during handling,
resuspend the sediment and remove the opaque supernatant before
sedimentation is complete. Repeat several times.
Section 4 : Column Packing
Nuvia ™ media can be packed using pressure, volumetric flow, or
vacuum packing methods. To pack columns for highly efficient
operation, it is recommended that a 20–50% slurry volume be used.
Packing Small Columns
This slurry packing method was designed to pack 25 ml of Nuvia
media in a conventional column with an internal diameter of 5–15 mm.
All buffers should be degassed. Because a relatively large volume of
slurry is required, it is recommended that a packing reservoir be used.
1. Prepare degassed 1.0 M NaCl, 20–50 mM buffer salt (see Table 2)
referred to herein as the packing buffer.
2. Nuvia media are shipped as a 50% slurry. Measure 50 ml of
suspended slurry into a 100 ml graduated cylinder. Allow the resin bed
to settle. Decant the shipping solution away from the resin bed.
3. Add 50 ml degassed packing buffer to the resin.
4. Seal the cylinder and rotate it to suspend the resin. Caution: Do not
mix with a magnetic stir bar as damage may occur. Larger amounts
of slurry may be mixed with a low-shear marine impeller at low to
moderate speed.
5. Add 10 ml packing buffer to the column. Pour in 75 ml resin slurry.
6. 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 0.1–1.0 cm.
7. Attach the column to your chromatography system, and purge the
column with starting buffer at linear velocities up to 600 cm/hr. If the
bed compresses,repeat steps 6 and 7.
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