affinity purification of receptor (was Re: avidin)

[Cornelius Krasel]

Bryan Crawford (bryanc at darwin.mbb.sfu.ca) wrote:
: This is sort of a tangent, but I wonder if anyone can suggest a method
: of eluting a biotinylated peptide from an avidin column which will not
: damage the column or the peptide?

You could try to elute with iminobiotin instead of biotin (has a lower
affinity to avidin than biotin). Maybe you can then regenerate your
column by extensive washing.

Another possibility (although I am not sure if it will work) is elution
with lipoic acid. Lipoic acid has been reported to elute successfully
so-called Strep-Tags (peptides which mimic biotin binding to streptavidin)
from streptavidin columns. See

Thomas G. M. Schmidt and Arne Skerra:
The random peptide library-assisted engineering of a C-terminal
affinity peptide, useful for the detection and purification of a
functional Ig Fv fragment.
Protein Engineering 6(1), 109--122, 1993.

--Cornelius.

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