UG-OPUS® 5-80R
OPUS® 5-80R Pre-packed Chromatography Columns
• If an OPUS® Column needs to be connected to an AKTA™ system which does not have TC
connections, the following item may be purchased from the GE Healthcare (GEHC) website
as a converter
o Part Number: 18116922
o Connector, 25mm TC – UNF 5/16” Female
o AKTA™ is a trademark of GE Healthcare
• Instructions for using a 3-way valve to connect the column inlet as well as purge air from the
inlet line can be found in Appendix 1
• Instructions on how to connect an OPUS® Column to chromatography skids can be found in
Appendix 2
1. Connect the column to the chromatography skid while limiting the entry of air to the inlet
port connection. Repligen suggests the following sequence for reducing air introduction to
the column:
1.1. The column may have off-gassed during shipment which leaves the ports dry; however,
the packed bed will remain hydrated in its storage solution. Top off the inlet and/or
outlet port with low salt equilibration buffer (e.g. 0.1M NaCl) prior to making
connections if needed.
1.2. With the outlet closed, hook up the column inlet under low flow (~50 cm/hr) with a Tri-
clamp (check Table 2 or 3 for the inlet and outlet size) so that no air is introduced into
the column.
1.3. Once the inlet has been connected under low flow, immediately stop the flow of liquid
into the column. Column outlet may now be opened via same instruction as Step 1.2.
1.3.1. If both the inlet and outlet port are open, the column may drain out and draw
air into the column flow path.
2. Connect outlet tubing to the outlet port and flush the storage solution from the column with
3-5 column volumes of RO/DI water or mild buffer (e.g. PBS).
Notes:
• When in contact with Ethanol solutions, high salt concentration buffers may precipitate
solids into the packed bed
• While flushing the Ethanol storage solution out of the column, high pressures are to be
expected due to the viscosity of the solution. Flow rate must be reduced if pressures
near >75% of the column packing pressure
• Observations of air exiting the column are common and will dissipate when the Ethanol
is fully cleared from the column
2.1. All the solutions loaded on to the column should be 0.22 or 0.45 µm filtered to reduce
column fouling.
2.2. Start storage solution flush with 50 cm/hr flow.
2.2.1. Flow rate may be increased as needed while maintaining conformance to the
resin supplier’s pressure recommendations. The column could yield high
pressure during Ethanol removal steps if flow rate is not monitored.
3. Equilibrate your column using 2-3 CVs of your equilibration process buffer or mobile phase.
4. To test for chromatographic performance and compare results to the CoA, a short
instruction guide can be found in Appendix 3.
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