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Bio-Rad Bio-Scale™ Mini UNOsphere SUPrA™ Affinity Cartridges User Manual

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Fig. 1. Flow performance of UNOsphere SUPrA™ media in Bio-Rad InPlace™ column
(20 cm x 20 cm) packed to 13% axial compression.

Section 3
Preparation for Packing

UNOsphere SUPrA affinity chromatography media are supplied fully hydrated in
20% ethanol as a 50% (v/v) slurry. For column packing, removal of the shipping
buffer is recommended. Small volumes of UNOsphere 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 with packing buffer is recommended.

Complete removal of fine particles from UNOsphere SUPrA media is not required
since the media are manufactured with a very narrow particle size range. If fine
particles (fines) have been generated during handling, resuspend (reslurry) the
media, let settle, then decant the supernatant containing the fines. Repeat this
process several times until a clear supernatant is obtained.

When preparing a homogenous slurry from settled material, take special care not to
crush the settled media with a mixing paddle; this can create fines. Use a side-to-side
motion or J-stroke with a

PTFE mixing paddle or other plastic paddle to disturb the

top layers of the settled bed until the slurry becomes homogenous. Alternatively,
gently roll the sealed container back and forth in a rocking motion to resuspend the
media.

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Linear flow (cm/hr)

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