GE Filter Kit HR 10 User manual

GE Healthcare
Life Sciences
Instructions 52-1777-00 AM Filter Kit HR10/HR16
Filter Kit HR 10, Filter Kit HR 16
These instructions describe how to change filters and O-rings on
HR 10 and HR 16 columns.

Table of Contents
31 Filter kit contents .......................................................................................
32 When to change the filter .....................................................................
33 How to change the filter ........................................................................
64 Troubleshooting .........................................................................................
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1 Filter kit contents
These filter kits (Code Nos. 18-3575-01/18-3585-01 respectively)
contain 10 filters and 4 O-rings.
2 When to change the filter
Change the filter on the top of your column when you observe an
increased backpressure, a space which has become visible between
adapter and filter, a loss of resolution and always before you perform
a thorough column cleaning.
Change the filter only at the top since this normally restores perfor-
mance and since changing the bottom filter may lead to loss of
efficiency.
3 How to change the filter
To change the filter you need a Filter tool, Code No. 18-3590-01,
consisting of a hook and a plunger.
ActionStep
Start the pump and pump buffer solution, e.g., the buffer
previously used, through the column at a flow rate of
1 to 1.5 ml/min.
Note:
Do not exceed recommended maximum backpressure.
1
Gradually rotate the red adjusting ring counter-clockwise.
Unscrew the red ring completely, and stop the pump.
2
Pull the adapter out by the red ring. Disconnect the column
top inlet tubing from the valve.
3
Check that some buffer remains over the chromatography
medium bed.
4
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ActionStep
Carefully and slowly bring up the loose
filter by using the hook (part of the Filter
tool). Do not press or puncture the medi-
um bed.
Note:
Only a very small amount of medium
should be in suspension after removing the
filter.
5
Check that the surface of the medium bed
is horizontal and even. If not, use the
plunger (part of the Filter tool) and careful-
ly stir up 2–3 mm of medium by rotating
it on the very top of the medium bed.
6
Wet a new filter in 20% ethanol. Put the
filter into the column, avoiding air bubbles
under it. Push it gently down to the medi-
um surface with the plunger. For HR 16
columns, use the wider end of the plunger.
Note:
As the filter is inserted, any medium which
has been stirred up will be packed onto the
top of the bed.
7
Make sure the filter is horizontal, and remove the plunger.8
Remove the liquid and traces of medium above the filter with
a pasteur-pipette. Rinse the walls and the top of the filter with
small aliquots of buffer until all the medium particles above
the filter have been removed.
9
Fill the volume above the filter with buffer.10
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ActionStep
Check the O-ring in the adapter. Change it if it is damaged or
asymmetrical. Do not use a sharp tool to remove the O-ring,
as this could damage the adapter and cause leakage.
11
Insert the adapter, aligning it so that it engages the slots in
the end piece.
12
Adjust the adapter to the medium bed by clockwise rotation
of the red adjusting ring until there is no space visible between
the adapter and the filter. Check that no large air bubbles
have been trapped between the medium and the adapter. If
a large air bubble has become trapped, carefully remove and
replace the adapter.
13
Connect the column inlet tubing to the valve.14
Start the pump and pump buffer at a flow rate of 1 to 1.5
ml/min for 5 to 10 minutes to pack the top of the medium
bed.
15
Stop the pump and disconnect the column inlet from the
valve.
16
Adjust the adapter to the filter again by clockwise rotation of
the red adjusting ring. When there is no space between the
adapter and the filter, rotate the red ring an additional half
turn for final positioning of the adapter.
17
Reconnect the column inlet to the valve.18
Test the column performance with a suitable sample.19
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4 Troubleshooting
Try the following if column performance is not restored:
•Wash the column according to the procedures described in the
“Instructions” supplied with each column.
•Repeat the procedure described with a new filter.
•If there is still no improvement, re-suspend 2–3 mm of the top of
the medium bed using the plunger as described above (Step 6).
Remove the suspension and replace the filter and adapter.
•As the final attempt to restore performance, check carefully that
the adapter is properly adjusted to the medium bed, and reverse
the flow direction in the column. Use a flow rate of 1 to 1.5 ml/min
for 10 to 30 minutes.
For further information contact your local GE Healthcare represen-
tative.
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For local office contact information, visit
www.gelifesciences.com/contact
GE Healthcare Bio-Sciences AB
Björkgatan 30
751 84 Uppsala
Sweden
www.gelifesciences.com/protein-purification
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© 2002-2012 General Electric Company – All rights reserved.
First published Sep. 2002
All goods and services are sold subject to the terms and conditions of sale of the company within
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Contact your local GE Healthcare representative for the most current information.
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52-1777-00 AM 08/2012
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