cytiva Filter Kit HR 10 User manual

Filter Kit HR 10, Filter Kit HR 16
Instructions for Use
These instructions describe how to change filters and O-rings on HR 10 and
HR 16 columns.
cytiva.com 52177700 AN

Table of Contents
1Filter kit contents ................................................................................... 3
2 When to change the filter .................................................................... 3
3 How to change the filter ....................................................................... 3
4 Troubleshooting ..................................................................................... 7
2 52177700 AN

1 Filter kit contents
These filter kits (Code Nos. 18357501/18358501 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
performance 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. 18359001,
consisting of a hook and a plunger.
Step Action
1Start the pump and pump buffer solution, e.g., the
buffer previously used, through the column at a flow
rate of 1 to 1.5 to ml/min.
Note:
Do not exceed recommended maximum
backpressure.
2Gradually rotate the red adjusting ring counter-
clockwise. Unscrew the red ring completely, and stop
the pump.
52177700 AN 3

Step Action
3Pull the adapter out by the red ring. Disconnect the
column top inlet tubing from the valve.
4Check that some buffer remains over the
chromatography medium bed.
5Carefully and slowly bring up the loose
filter by using the hook (part of the Filter
tool). Do not press or puncture the
medium bed.
Note:
Only a very small amount of medium
should be in suspension after removing the
filter.
6Check that the surface of the medium bed
is horizontal and even. If not, use the
plunger (part of the Filter tool) and
carefully stir up 2–3 mm of medium by
rotating it on the very top of the medium
bed.
4 52177700 AN

Step Action
7Wet a new filter in 20% ethanol. Put
the filter into the column, avoiding air
bubbles under it. Push it gently down
to the medium 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.
8Make sure the filter is horizontal, and remove the
plunger.
9Remove 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.
10 Fill the volume above the filter with buffer.
11 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.
12 Insert the adapter, aligning it so that it engages the
slots in the end piece.
52177700 AN 5

Step Action
13 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.
14 Connect the column inlet tubing to the valve.
15 Start the pump and pump buffer at a flow rate of 1 to
1.5 to ml/min for 5 to 10 to minutes to pack the top of
the medium bed.
16 Stop the pump and disconnect the column inlet from
the valve.
17 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.
18 Reconnect the column inlet to the valve.
19 Test the column performance with a suitable sample.
6 52177700 AN

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 to ml/min for 10 to 30 to minutes.
For further information contact your local Cytiva
representative.
52177700 AN 7

cytiva.com/protein-purification
Cytiva and the Drop logo are trademarks of Global Life Sciences IP Holdco LLC or
an affiliate.
All other third-party trademarks are the property of their respective owners.
© 2020 Cytiva – All rights reserved.
All goods and services are sold subject to the terms and conditions of sale of the
supplying company operating within the Cytiva business. A copy of those terms
and conditions is available on request. Contact your local Cytiva representative for
the most current information.
For local office contact information, visit cytiva.com/contact
52177700 AN V:5 07/2020