RE: The best way to separate glycoproteins

Patrick McAtee (PatrickM@genelabs.com)
Wed, 11 Feb 1998 17:09:22 -0800

There's a better way if the resin still exists: Use Phenyl boronate
from Amicon, high octane version usually works for protein (higher
density load, they make three types of resin). Equilibrate in HEPES at
neutral to slightly basic pH and bump with Tris or pH adjust. You can
try sugars to elute but addition of Tris does the trick. I did some
GM-CSF from yeast broth this way and the results were quite pretty. My
process for GM-CSF was certainly alot better than what was given to me
(multiple HPLC steps in weird stinky buffers resulting in "brown"
cytokines) by a company from the land of endless rain. (Of course I'm in
Northern California, I guess I can't talk)

C. Patrick McAtee, Ph.D.
Group Leader,
Microanalytical Chemistry (it keeps changing names !)
Director of Mass Spectrometry Laboratory
Genelabs Technologies, Inc.
> ----------
> From: ccarrawa@mednet.med.miami.edu
> Sent: Tuesday, February 10, 1998 4:58 AM
> To: Recipients of ABRF List
> Subject: Re: The best way to separate glycoproteins
>
>
> Lectin-agarose columns are great for this purpose. The
> appropriate
> lectin must be selected for each glycosylated protein, which can
> be
> done by screening on blots. Common lectins used for this purpose
>
> include Con A and wheat germ agglutinin, but others are also
> used.
>
> Another consideration is release of the bound glycoprotein with
> the
> appropriate hapten sugar. It is not always best to select the
> lectin
> with the highest avidity of binding, since it will be more
> difficult
> to release the glyprotein.
>
> Good luck!
>
>
> ______________________________ Reply Separator
> _________________________________
> Subject: The best way to separate glycoproteins
> Author: Bao-Shiang Lee <boblee@uic.edu> at SMTPMED
> Date: 2/10/98 7:25 AM
>
>
> Hi, everyone, I need some suggestions in separating glycoproteins from
>
> non-glycoproteins. Are there columns that can do the job. I would like
> to
> also separate glycoproteins. Best Wishes. Bob
>
>