Re: HPLC: Boronate Affinity Chromatography of Glycopeptides
Dan Crimmins (crimmins@pathbox.wustl.edu)
Tue, 16 Mar 1999 15:08:14 -0600
At 09:15 AM 3/16/99 -0800, Apffel, Alex wrote:
>We are trying to use boronate affinity columns from TosoHaas to isolate
>glycosylated recombinant proteins and glycosylated peptide fragments
>produced upon proteolytic digestion of the recombinant glycoproteins. These
>are primarily, but not exclusively N-linked glycosylation. As an
>alternative to Lectin Affinity chromatography, the use of the boronate
>columns sounds attractive because we could simultaneously isolate N- and
>O-linked glycopeptides from non-glycosylated peptides and elute the
>fractions in mobile phases that are relatively friendly to subsequent LC/MS
>and CE/MS analysis. A suggestion has been made that boronate columns work
>well for glyco-amino acids, less well for glycopeptides and poorly for
>glycoproteins. Can you shed some light on this for us.
>
>Alex Apffel, Ph.D.
>Biomeasurements Group
>Hewlett-Packard Laboratories
>3500 Deer Creek Road
>Palo Alto, CA 94304 USA
>Tel 650-857-6090
>Fax 650-852-8502
>email alex_apffel@hpl.hp.com
>
>
>
Alex,
Boronate affinity columns have been used to isolate glycated-Hb from
native Hb in clinical samples, so this procedure can be used for
glycoproteins.
Regards,
Dan L. Crimmins
Washington University School of Medicine
Dept. Pathology/Division of Laboratory Medicine
660 S. Euclid Ave., Box 8118
St. Louis, MO 63110
Phone: 314-454-8514; Fax: 314-454-5208
e-mail: crimmins@labmed.wustl.edu