I am working on nanobody based purification system and in order to reuse it, I have to separate the nanobody from its ligand.
I assume you have your antigen fished out by a NB-based affinity column, and now want to elute the antigen.
NB are a special form of antibodies, so try the methods that break the bond between antibodies and antigen (Gly/HCl pH 2, caotropic agents like urea, high salt…). Remember to replace that eluent by a more physiological buffer ASAP to avoid permanent damage. Harlow & Lane 1988 is still a good source of information, despite its age.
Engelbert Buxbaum thank you for your reply. I have a problem with this method as my NB is immobolized on a solid support by His tag-Ni2+ intereaction. When I apply a low pH buffer, it also breaks the immobilization bond.
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