Where I work we routinely analyze carbohydrates on glycoproteins. To get a sense
of the % glycosylation, you can just look at the apparent molecular weight on a
gel vs. the theoretical peptide weight for the protein backbone. To analyze
whether there are N or O-link structures (or both), digest with N-glycanase
(PNGase from Oxford glycosystems) and/or O-glycanase (many forms so check out
Oxford for this also) and run on a gel for apparent molecular weight. For more
detailed analysis (types of sugars, molar ratios, sialic acid content,
oligosaccharide mapping, etc..) you would need equipment designed for that
analysis. A common tool is the Dionex HPAE-PAD which can be used to quantify
sugars by type (mannose, galactose, sialic acids etc..). Alot of information
about carbohydrates can be gleaned from peptide digests run on LC/MS. But this
does not quantify the species.
Shawn Novick
Immunex Corp.
Seattle, WA.
_______________________________________________________________________________
Subject: carbohydrate quantitation
From: Kelly S Cass-Samodral <CASS-SAMODRAL_KELLY_S@lilly.com> at Internet
Date: 3/31/98 10:06 AM
Hello ABRF members,
I was asked today what is the method of choice for the quantitation of
glycoforms on a glycoprotein? Is this being done routinely, ie QC? Are there
other ways to monitor the same information?
All answers accepted. Thank you in advance for your time.
Kelly Cass-Samodral
KSCS@lilly.com
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Date: Tue, 31 Mar 1998 10:06:06 -0500
From: Kelly S Cass-Samodral <CASS-SAMODRAL_KELLY_S@lilly.com>
Subject: carbohydrate quantitation
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