breaking biotin-streptavidin interaction

U12201 at uicvm.uic.edu U12201 at uicvm.uic.edu
Wed Sep 27 17:09:47 EST 1995


In article <44a0pa$cl6 at rover.ucs.ualberta.ca>, doug at hobbes.chem.ualberta.ca (Doug Craig) says:
>
>Is there a relatively easy way to remove a biotin-labelled peptide from a
>streptavidin-labelled bead? I would prefer if the streptavidin wasn't
>killed by the process but if it is it doesn't really matter. It is
>imparative that the peptide survive and retain the biotin label. The peptide
>will be sufficiently small that its denaturation isn't a problem.
>
>Any suggestions?
>
>Thanks,
>
>Doug
>
>.............................................................................
>Doug Craig
>Chemistry
>University of Alberta
>Edmonton, AB T6G 2G2
>doug at hobbes.chem.ualberta.ca
>.............................................................................

Boy do I have an easy solution for you: Pierce Chemical sells
what is called "mono-meric avidin Sepharose" - when avidin
is monomerized it drops the affinity for biotin to a level
where it still binds at about 10e-08, but can easily be
displaced by either biotin or other chaotrophes. This way the
peptide is easily eluted (with biotin) which is then removed
by dialysis (use the Slid-A-Lyzer) for convenience.
Alternatively, you may want to use imino-biotin instead of
biotin for the peptide lable, then you can elute off immobilized
avidin by a pH drop, but this is a whole other story.
Keld Sorensen   KeldS at uic.edu



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