Skip to main content
. 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 3.

CIEF and microchip IEF (mIEF)]

Analyte Mode Capillary coating Capillary Detector Ampholyte Catholyte Anolyte Notes Ref
In-house EPO,
Fc-fusion protein, and IgG
iCIEF Fluorocarbon
(Protein Simple)
iCE280 Analyzer;
50 mm, 100 µm id
UV 280 nm Pharmalyte pH 3–10,
4–6.5, 5–8, and 8–10.5
0.1 M NaOH
in 0.1% MC
0.08 M
phosphoric acid
in 0.1% MC
Wide range of
Therapeutic protein
applications
[51]
In-house noninfectious
virus-like particles
iCIEF Fluorocarbon
(Protein Simple)
iCE280 Analyzer;
50 mm, 100 µm id
UV 280 nm Pharmalyte pH 2.5–5
and 3–10
0.1 M NaOH
in 0.1% MC
0.08 M
phosphoric acid
in 0.1% MC
Charge characterization
of virus-like particles
[52]
In-house vaccine carrier
protein
iCIEF Fluorocarbon
(Protein Simple)
iCE280 Analyzer;
50 mm, 100 µm id
UV 280 nm Pharmalyte pH 3–10
and 4–6.5
0.1 M NaOH
in 0.1% MC
0.08 M
phosphoric acid
in 0.1% MC
Characterization of
polysaccharide vaccine
carrier protein
[53]
In-house IgG2κ
mAb
CIEF PDMA 180 mm,
50 µm id
LIF ex. 543.5 nm /
em. 590 nm
Pharmalyte pH 3–10,
0.001% BSA
20 mM NaOH 20 mM
phosphoric acid
Determination of
deamidation rates in
mAb
[54]
Trypsinogen,
β-lactoglobulin, BSA,
and ovalbumin
CIEF 4% acrylamide
0.6% cross-
linker
Various capillary
lengths and id
LIF ex. 488 nm BioRad pH 3–10 20 mM NaOH 10 mM
phosphoric acid
Microchip interface for
2D CIEF and CGE
separations
[55]
β-lactoglobulin A,
hemoglobin A, myoglobin,
α-chymotrypsinogen A,
ribonulcease A,
cytochrome c, lysozyme
CIEF Seven PEEK tubing
segments, 1.55 cm,
395 µm id connected
by Nafion membrane
0.2 cm, 330 µm id
TOF and Orbitrap Various MS-compatable
carrier ampholytes
Various electrolytes at each
Nafion junction modified the local
pH of
the carrier ampholyte
Segmented capillary for
selective mobilization
[60]
Insulin receptor and
protein tryptic digests
CIEF LPA 50 cm, 50 µm id Sheath flow ESI-
Orbitrap-MS
Glutamate, asparagine,
glycine, proline,
histidine, and lysine
0.1% Formic
acid, pH 2.5
0.3% ammonium
hydroxide, pH 11
Amino acids used as low
MW ampholyte
[64]
In-house IgG
mAb
iCIEF Proprietary
photoreactive
layer
12-channel cartidge;
50 mm, 100 µm id
Chemiluminescence
from secondary
antibody
Pharmalyte pH 5–8
(30%) and
pH 8–10.5 (70%)
0.1 M NaOH
in 0.1% MC
0.08 M
phosphoric
acid in 0.1% MC
CIEF with immunoassay
detection
[65]
Donated mAb products imIEF Uncoated quartz MCE-2010 system,
2.7 cm
UV 280 nm Protein Simple pH 3–10,
5–8, and 8–10.5
300 mM NaOH,
0.4% HPMC
200 mM
phosphoric acid,
0.4 % HPMC
mIEF of mAb charge
variants
[141]
Model Proteins iMIF 100 µm id Immunoblot Polyprotic carboxylic
amino acids
20 mM lysine 20 mM arginine Integrated microchip for
separation and
immunoblot
[142]

Capillary: actual length, inner diameter. Bovine serum albumin (BSA), Linear polyacrylamide (LPA), Methylcellulose (MC), Polydimethylacrylamide (PDMA), Polyvinyl alcohol (PVA),