Protein Glycosylation: Implications for In Vivo Functions and Therapeutic Applications - Advances in Biochemical Engineering Biotechnology - page 190

Table 4 (continued)

Analysis Techniques followed Remarks Ref.

[e.g. monosaccharide residue, anomericity ( ) or

linkage, absolute stereoisomer( D / L ) of monosaccharide]
except substitution pattern. Mono-saccharides are
analyzed by normal phase HPLC or MALDI-MS

a / b

2. NMR spectroscopy Potentially the most informative for glycan [272]

structure, linkage position and anomeric configuration
analysis. Relatively insensitive

3. Gas chromatography (GC)/MS Composition and linkage of monosaccharides are [257]

best determined after methanolysis

4. Fast atom bombardment(FAB) Sequence, branching and more linkage information [273]

or liquid secondary ion(LSI) MS can be obtained. Moderate sensitivity. Sample

derivatization necessary

5. Tandem MS (MS-MS) Sequence, branching pattern and some linkage [274]

information can be obtained.
O -linked glycosylation also analyzed

6. Electronspray ionization(ESI) MS Useful in quality control for recombinant proteins and site [275]

analysis of tryptic glycopeptides. Readily interfaced with
HPLC or CE systems. It can differentiate between
O - and N -linked glycans, and also between complex,
hybrid or high mannose forms

7. MALDI TOF-MS Simple, quick and quite insensitive to salts or detergents. [276]

Useful for detailed structural analysis including linkage
and arm specificity. Unlabelled glycans can be analyzed.
Derivatization increases detection limits.