affinity purification of receptor (was Re: avidin)

Cornelius Krasel krasel at alf.biochem.mpg.de
Mon Aug 29 13:15:20 EST 1994


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.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795 */
/* "People are DNA's way of making more DNA." (Edward O. Wilson, 1975)    */



More information about the Methods mailing list