Cleaning-in-place (CIP)
O
ne of the most important aspects of production chromatography is the maintenance
of the packed column by cleaning-in-place (CIP) procedures. Regular CIP of the packed
column between production batches is needed to ensure the proper product quality
and the expected media and equipment life. CIP removes precipitated material,
strongly bound substances, and other contaminants from the column bed without
dismantling the column.
Recommended CIP procedure
CIP protocols should be included as part of the general process. CIP of the media
should take place when required, for example after each production batch, when an
unusually high back-pressure is generated over the column, or when a change in peak
symmetry or discoloration of the media is observed. See the recommended cleaning
procedures in the table below.
Media Contaminants Suggested procedure Follow-up
Sepharose
F
ast Flow and
Sepharose
High Perform-
ance based
media
Soluble proteins/
electrostatic
interaction.
Wash with 0.5 column volumes of
filtered 2 M NaCl for a contact time
10 to 15 minutes.
Regenerate to start
conditions. See
instructions for
respective media.
Soluble proteins/
h
ydrophobic
interaction.
Wash with 2 to 3 column volumes of
f
iltered low ionic strength buffer or
water for a contact time of 10 to 15
minutes, or
wash with 1 column volume of 30%
isopropanol for a contact time of
10 to 15 minutes, or
wash with 2 to 4 column volumes of
0.5 to 1 M NaOH for a contact time of
1 to 2 hours.
Regenerate to start
c
onditions. See
instructions for
respective media.
Precipitated
p
roteins
Wash with 2 to 4 column volumes of 1
M NaOH for a contact time of 1 to 2
hours. Use reversed flow, or
wash with 2 to 4 column volumes of a
mixture of 1 M NaCl/1 M NaOH for a
contact time of 1 to 2hours. Use
reversed flow.
If necessary, wash
with 1.5 column
volumes of water
followed by 1
column volume of 2
M NaCl (not
HIC).Regenerate to
start conditions,
see instructions for
respective media.
Regenerate to start
conditions, see
instructions for
respective media.
6 Maintenance
6
.7 Cleaning
116
BPG Columns Operating Instructions 29193223 AB
To Next Page
Loading...
