> ----------
> From: Michael N. Horst, Ph.D.
> Sent: Thursday, February 12, 1998 5:11 AM
> To: Patrick McAtee
> Subject: Re: The best way to separate glycoproteins
>
> Patrick- I was intrigued with your post. I recall similar techniques
> from
> years ago, but I have lost touch with the current versions. So.... Do
> you
> have references to the method ? Thanks for making this info available
> to the
> group. This is why it is so worthwhile. Mike.
> ******************
> Michael N. Horst
> Mercer Medical School
> Macon GA 31207
> ******************
>
> Patrick McAtee wrote:
>
> > 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
> > >
> > >
>
>
>