Biomedical Engineering Reference
In-Depth Information
Table 4
(continued)
Analysis
Techniques followed
Remarks
Ref.
Enzymatic release (N-glycans)
1. PNGase F
Cleaves
b
-aspartylglycosylamine bond of all classes of glycans
[258,
2. PNGase A
Cleaves sugars having
a
1,3-linked core fucose,
259]
resistant to PNGase F
3. Endo H
Selectively cleaves oligomannose and hybride type
[259,
structure between the two Gn residues within the core
260]
4. Endo D
Cleaves all classes of
N
-glycans between the two Gn residues
[259]
within the core
5. Endo F
1
,F
2
,F
3
Cleave within the chitobiose core, but Endo F
1
digests only
[259]
oligomannose and hybrid structures, Endo F
2
digests oligomannose
and biantennary structure and Endo F
3
is specific for bi- and
triantennary glycans.
B) Labelling of glycans
Radioactive labelling
Classical method, sensitivity of
at Reducing termini
detection increased to femtomole.
Tritium
Largely replaced by fluorescent compound labelling
Fluorescently labelled
Sensitivity of detection increased and ease in
separation due to introduced charge
1. 2-aminobenzamide (2-AB)
HPLC, reversed phase (RP)-HPLC, weak anion exchange,
[261]
high pH anion exchange, BioGel P4 gel permeation
chromatography, matrix assisted laser desorption ionisation
(MALDI) mass spectrometry (MS) and electron spray
(ES) MS could be deployed to separate glycans
2. 2-anthranilic acid (2-AA)
Electrophoretic separation of glycans by SDS-PAGE
[262]
3. 8-aminonapthalene-1,3,6-
Particularly useful for electrophoretic separation
[262]
trisulphonic acid (ANTS)
of neutral sugars, allowing resolution from acidic glycans
4. 2-aminopyridine (2-AP)
Used extensively in RP-HPLC and High performance
[263]
capillary electrophoresis (CE)
5. 2-aminoacridone (AMAC)
Suitable for SDS-PAGE of glycans and micellar electrokinetic
[264]
capillary chromatography (MECC).Also compatible with
RP-HPLC and MALDI time-of-flight (TOF) or nanolitre flow ES-MS
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