Affinity chromatography: How to pack columns / cartridges

Описание: Let us show you how to pack an empty cartridge / column with one of our protein purification resins.

The column / cartridge Assembly Press can be found here:
https://cube-biotech.com/products/pro...

The example resin that is used in this video is our in-house developed Ni-INDIGO agarose resin:
https://cube-biotech.com/products/pro...

The column / catridge that has been used can be found here:
https://cube-biotech.com/products/pla...

Individual steps of the packing protocol:
0:00 - Intro
0:11 - Required equipment
0:46 - Adding the first frit
0:59 - Mix resin with storage buffer
1:08 - Fill syringe with storage buffer
1:17 - Attach empty tube to syringe
1:23 - Fill tube with storage buffer
1:30 - (1) Add purification resin
1:39 - Remove most of the storage buffer
1:47 - (2) Add purification resin
1:58 - Top up tube with storage buffer
2:11 - Column Assembly
2:19 - Add 2nd frit
2:26 - Add sealing plug
2:36 - Apply pressure
2:51 - Add sealing sleeve
3:01 - Add top stop plug
3:05 - Endcard

The full protocol can be found here (Also as PDF-file):
https://cube-biotech.com/packing-of-c...
or down below in this description.

If you have more questions regarding this protocol or our products in general, feel free to write an email to "[email protected]" or simply use the Chatbot at https://cube-biotech.com/ to get live support.

Cube Biotech is an expert in all matters of protein purification. We use all of our products in-house and are also active in R&D in the field of protein purification!

Full Protocol:

Part 1 - Column Preparation:
Step 1: Push the smaller frit into the empty tube.

Step 2: Shake the resin bottle to mix beads with storage buffer. This has to be done because otherwise the beads settle on the bottle bottom.

Step 3: Fill a syringe with resin storage buffer and ensure that no air is left in the syringe.
Note - All steps up to step 9 from now on: It is essential to avoid any air bubbles in the purification cartridge / tube from now on.

Step 4: Attach the empty tube to the syringe

Step 5: Fill approx. one quarter of the empty tube with agarose storage buffer. Again, ensure that no air bubbles are in the buffer.

Step 6: Add 500 µl mixed purification resin to the column. Make sure that the resin slurry is still properly mixed and the beads did not settle.

Step 7: Remove most of the storage buffer through the use of the syringe. Do not leave the resin beads dry! Otherwise air bubbles might form in between the small agarose beads.

Step 8: Add another 500 µl of purification resin. Again ensure that it is still properly mixed.

Step 9: Top up the tube with resin storage buffer. Up until it starts to flow over. No possible space for air bubbles should remain.


Part 2 - Column Assembly
Step 10: Add top frit. It is normal that some of the just added storage buffer spills over here. This is actually a good sign because it means no air was left in the tube.

Step 11: Add the sealing plug and place the tube in the cartridge holder.

Step 12: Apply pressure on the cartridge to seal the resin airtight between the two frits. Again it can occur that some form of liquid spills over when the pressure is added.

Step 13: Screw on the bottom stop plug and add the sealing sleeve.

Step 14: Complete column by adding the top stop plug.


Agarose storage Bufffer composition:
20% ethanol
20 mM sodium acetateLet us show you how pack an empty cartridge / column with one of our protein purification resins.