p. 10
5. Cleaning, Sanitization and Storage
For best performa nce of coupled CNBr-act ivated Sepharos e 4 Fast Flow over
a long working life, follo w the general proce dures described b elow. In all
cases, we recommend tes ting the procedu res at small scale f irst.

Equilibration

After packing, and before a chromatographic run, equilibrate with working
buffer by washing wit h at least 5 bed vol umes.

Cleaning-In-Place

Cleaning-in-plac e, (CIP), is a cleani ng procedure which re moves
contaminants such as li pids, precipita tes or denatured pro teins that may
remain in the packed colu mn after regener ation. Such contam ination is
especially likely whe n working with cr ude materials. R egular CIP prevent s
the build-up of these co ntaminants in th e packed bed, and hel ps to maintain
the capacity, flow pro perties and ge neral perform ance of the medium.
A specific CIP protocol s hould be designe d for each process acco rding to the
type of contaminants pre sent and stabili ty of coupled ligand . The frequency
of CIP depends on the nature a nd the condition o f the starting m aterial and
other process requirem ents, but one CIP cy cle is generally reco mmended
every 1–5 separation cycle s. Following are ge nerally recommen ded
procedures.

CIP protocol

Precipitated or Wash wi th 2 column volume s of
denatured 6 M guanidine hyd rochloride. Wash
substances substances immediately with at least
5 column volumes of sterile fi ltered binding
buffer.
Hydrophobically Wash the column with 2 colu mn volumes of
bound substances a non-ionic detergen t (conc. 0.1–0.5%).
Wash immediately with at leas t 5 column
volumes of sterile filtere d binding buffer.