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· Determination of non-human sialylation by quantitative determination of NeuAc:
NeuGc ratio using LudgerTag DMB sialic acid analysis kit and LudgerSep R1
HPLC with fluorescence detection) |
 |
· Monosaccharide composition analysis using either fluorescence HPLC or HPAE-PAD
(high pH anion exchange with pulse amperometric detection) |
· Release of N- and O-linked glycans from the glycoprotein or glycopeptides by :
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- a. Endoglycosidases (for N-linked glycans only) |
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- b. Hydrazinolysis (in N+O mode, N-mode or O-mode) |
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- c. Ludger LiberateTM glycan release system in N+O mode or O-mode |
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· Fluorescent labeling of released glycans prior to chromatography or mass
spectrometry using |
·one of the following LudgerTag labeling systems :
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- a. LudgerTag 2-AB (2-aminobenzamide) for HPLC and MS |
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- b. LudgerTag 2-AA (2-aminobenzoic acid) for HPLC, MS and carbohydrate gel
electrophoresis |
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- c. LudgerTag AA-Ac for HPLC, MS and capillary electrophoresis |
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· Oligosaccharide mapping of fluorescently labelled glycans by :
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- a. LudgerSep Cx anion exchange HPLC (for glycan charge analysis) |
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- b. LudgerSep N1 amide HPLC (for analysis of charged and neutral glycans) |
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- c. Dionex HPAE-FD (fluorescence mode high pH anion exchange used as
an orthogonal separation to complement the N1 profile) |
|
| · Oligosaccharide mapping of non-labelled glycans by HPAE-PAD |
· Mass spectrometric analysis (including MSn fragmentation analysis) of derivatized
and native glycans and glycopeptides by ion trap MS (Shimadzu Axima QIT) |
· Glycan sequencing using exoglycosidase digestions in combination with HPLC and
MS analysis |
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