Antibody Affinity Columns

Dayanidhi Raman maraman at ksu.ksu.edu
Thu Apr 13 10:05:17 EST 1995


lees at acpub.duke.edu (Steve Lee) writes:

>I was wondering if anyone knew of a good "regeneration buffer" for
>antibody affinity columns... comercial affinity column kits always come
>with a "regeneration buffer" to keep binding capacity high..  if anyone
>knows of one, please mail me..

>and if anyone has ever had trouble with albumin contamination in
>purifying blood proteins from plasma using an affinity column please
>mail me as well..

>thanks alot!

>steve
Hi, I would you recombinant Protein G sepharose to make antibody affinity column in your case because the albumin binding site in Protein G is deleted in it. If you still have some albumin you might try anti-albumin affinity chromatography to get rid off albumin but I haven't done this & so I am not sure whether it is practical to do it.  
Regarding regeneration, you can wash with acidic and alkaline buffers 3x alternatively. 
Good luck.

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