Separation of tryptic digest
sstriepk at CRL.COM
Sun Oct 27 23:39:34 EST 1996
Boronic acid gel, which is sold through Aldrich and Sigma, might do the
job. The gel binds carbohydrates in the same manner as borate. Adsorption
and elution from the gel is controlled by pH; neutral or slightly basic
to bind the sugar or monosaccharide, acid pH to elute. Aldrich has a good
application sheet for the material along with a list of references.
You might also want to consider using enzymes other than PNGase F to
study the glycopeptides. There might be considerable heterogeneity of the
glycosylation of the peptides that could be reduced by digestion with
NANase (neuraminadase) and other exoglycosidases.
On 27 Oct 1996, Martin Kaliwoda wrote:
> Dear Glycoscientists!
> I am searching for an easy (like always) method to separate
> peptides from glycopeptides.
> After a tryptic digest and RP chromatography, I would like to identify
> glycopeptides. Because of the complex peak pattern of the RP map
> it is unpossible to use a PNGase digest after the tryptic digest
> and compare or fractionate the RP maps.
> A possibility would be the use of a ConA column, but I think that the
> ConA material is too specific.
> I would prefer a HPLC method, because there is only a mg amount of material.
> So if anyone of you has suggestions or experience with this problem, please
> post to the newsgroup, or mail to e8327091 at stud1.tuwien.ac.at.
> Thanks a lot
> Martin Kaliwoda
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