My question is simple:
How much worse is it to do IMAC puricifation without a column?
My plan is to just mix the resin with and E.Coli lysate, wash and elute it with centrifugation steps.
I realize that the consentration of the eluate will be lower at least, but for my application I don't care much about consentration or yield.
I'm just wondering if there is something I'm missing here?
Sincerely
Markku Syrjänpää
Batch or column purification?
Ni-NTA Agarose Resin is optimal for affinity purification of His-tagged fusion proteins by batch or column purification
original text
https://www.serva.de/www_root/documents/42139%20SERVA%20Ni-NTA%20Agarose%20Resin%20Ver%200215_e.pdf
http://wolfson.huji.ac.il/expression/local/Ni_NTA_Hi_Bind_Resins.pdf
@Markku You can do this, it works like the procedure woith magnetic bead. But take care not to mechanically destroy the particles by high g values. You can find a video of the magbead procedure here, it works nearly similar. https://www.youtube.com/watch?v=4JBWN5G8EBg
If you elute your target protein by just adding elution buffer, you might lose some of your material, as it will be hidden in the void volume of the resin.
If possible, re-use some emptied, old, spin column as carrier for the NTA resin, then you may centrifuge your samples for quantitative recovery.
As Antinia already pointed out. Batchspin is a good option here.
How this works can be seen here for example:
https://www.youtube.com/watch?v=9kMMicQfgJM
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