UG-OPUS® 5-80R
16
OPUS® 5-80R Pre-packed Chromatography Columns
7.
Introduce flow to the column at a low flow rate (i.e. 50 cm/hr) to flush trapped air from the
column outlet.
In the absence of a 3-way valve:
1.
A “T” line can be connected between the column and chromatography system.
2.
The “T” line can be used as described above for purging air.
3.
After air has been purged, the purge line can be clamped or closed with a stopper.
4.
The column outlet can then be connected to the chromatography system for normal use.
6.2
Appendix 2: Connecting an OPUS® column to Chromatography Skids
1.
Connect the inlet of the column to the chromatography system tubing set. At this point, the
outlet of the column should be closed and disconnected.
Notes:
•
Use of stainless steel valves connected directly to the inlet and/or the outlet ports is not
recommended. The additional weight to the top of the column will increase the risk of
damage to the column hardware
2.
Start flowing the mobile phase through the system at <50% of the recommended operating
flow rate for your chromatography resin/process. During this operation, the flow will be
split. One portion of the mobile phase will enter the bypass line and other portion will enter
the inlet line.
Notes:
•
The mobile phase will not enter the column because the column outlet is closed (with
the pinch valve, or outlet cap) thus creating a stop barrier for the flow. The fraction of
the flow that enters the inlet line will dislodge the trapped air in the tube and connector
3.
With the flow split to the bypass and inlet lines, air bubbles will begin to travel upward in the
inlet tube and will be evacuated through the column bypass line into the system outlet. To
ensure all the air is evacuated, tap and/or shake the inlet tube and inlet connector.
4.
After all the air has been removed from the column inlet and connector, close the bypass
line pinch valve and connect the outlet line to the column with the outlet pinch valve in the
open position.
5.
Run mobile phase through the column to purge the outlet of air.
6.3
Appendix 3: Column Performance Testing
Follow the steps below to measure the plate count and asymmetry of your OPUS® Column. Please
note, minor differences (plus or minus 10-20%) in the measured plate count and asymmetry noted
on the column CoA or CoQ are to be expected. Sources of variation include:
•
Chromatography instruments for measurement
•
Chromatography system
•
Operator variability
•
Normal variability within the test methods
o
Flow rate
o
Sample volumes
o
Equilibration/plug solutions
o
Injection method
If the plate count and asymmetry measurements are within defined acceptance limits (reference the
column’s CoA or CoQ “QC Release Data”) then the column should be considered fit for purpose.
1.
Remove column storage solution.