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. Author manuscript; available in PMC: 2015 Jul 8.
Published in final edited form as: Anal Methods. 2014 May 13;6(15):5427–5449. doi: 10.1039/C4AY00447G

Table 6.

CE-based analysis of protein glycosylation

Analyte Mode Capillary BGE Detection Label Notes Ref.
N-glycans of mAb1 CGE 48 capillary array,
50 cm each
Applied Biosystems
Pop-7™
LIF ex. 473 nm /
em. 520 nm
APTS High-throughput glycan analysis [112]
(1) Neu5Gc and
(2) α1,3-Gal
ontaining N-glycans
Partial-filling CE 40 (30) cm,
50 µm id
DB-1 capillary
100 mM Tris-acetic acid, 0.05%
HPC, pH 7.0
LIF ex. 488 nm /
em. 520 nm
APTS Reaction with (1) anti-Neu5Gc or
(2) α-galactosidase inject prior to
sample
[115]
Non-human
-glycans
CGE 40 (30) cm,
100 µm id
DB-1 capillary
100 mM Tris-borate,
5% PEG, pH 8.3
LIF ex. 325 nm /
em. 405 nm
2-AA Analysis of commercially available
mAbs
[116]
α1,3-Gal containing
N-glycans
CGE 60 (50) cm,
50 µm id
eCAP NCHO coated
Beckman Coulter Carbohydrate
Separation Gel Buffer-N
LIF ex. 488 nm /
em. 520 nm
APTS and
AMAC
Ultrasensitive detection method [117]
N-glycans of mAb CGE 50 (40) cm,
50 µm id
PVA coated
Beckman Coulter Carbohydrate
Separation Buffer
Or 40 mM EACA-acetate,
0.2% HPMC
LIF ex. 488 nm /
em. 520 nm
APTS Glycans of mAbs from NS0 cells [118]
N-glycans of fusion
protein
CE 90, 60, or 43 cm 0.7 M ammonia and 0.1 M
EACA in 70% MeOH
TOF-MS APTS Alkaline CE-MS method [119]
N-glycans (1) CGE
(2) CE
Various capillary
lengths and coatings
(1) Beckman glycan separation
buffer or POP-7 polymer
(2) 40 mM EACA, 131 mM
acetic acid, pH 4
(1) LIF ex. 488
nm /
em. 512 nm
(2) TOF-MS
APTS CGE-LIF and CE-MS methods
compared to the CE-MS method in
ref [119]
[120]
N-glycans of mAbs CE 50 (40) cm,
50 µm id
N-CHO coated
Beckman Coulter Carbohydrate
Separation Gel Buffer
LIF ex. 488 nm /
em. 520 nm
APTS CE-LIF as an orthogonal technique
to MS
[121]

Capillary: Actual length (effective length), inner diameter, coating

2-aminoacridone (AMAC), 2-aminobenzoic acid (2-AA), 9-aminopyrene-1,3,6-trisulfonic acid (APTS), ε-aminocaproic acid (EACA), Galα1–3Gal (α1,3-Gal), Hydroxypropyl cellulose (HPC), Hydroxypropylmethylcellulose (HPMC), N-Glycolylneuraminic acid (Neu5Gc), Polyvinyl alcohol (PVA), Poly(ethylene glycol) (PEG),