Capillary zone
electrophoresis (CZE):
High field strengths are used to separate proteins introduced into the
capillary by either electrokinetic or hydrodynamic means. Differences
in charge-to-mass ratios of a heterogeneous solution lead to discrete
zones which appear as peaks on the electropherogram. This free solution
technique requires only minute amounts of sample, and multiple runs can
easily be automated. Untreated fused silica capillaries used at
physiological pH can produce fast separations (a few minutes) due to
electroosmotic flow (EOF). Coated capillaries are used for more basic
proteins to prevent adsorption on the capillary wall.
Capillary gel electrophoresis (CGE):
A UV transparent linear polymer is used as a molecular sieve analogous
to traditional SDS-PAGE. Separation is based solely on size. Molecular
weight determinations are made by comparison to standards.
Capillary isoelectric focusing (CIEF):
As with traditional isoelectric focusing, a pH gradient is created
using ampholytes. Samples will migrate through this gradient until
their net charge becomes zero. Once the sample components are separated
into bands based on their prospective pIs, a low pressure is applied to
move them past the detector window.
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Micellar
electrokinetic capillary chromatography (MECC):
Micelles formed with traditional surfactants are used to aid in
separations. Cationic sample components are electrostatically attracted
by these micelles and elute last as the EOF works against micelle
migration. Migration of anionic components is faster than migration of
neutral molecules, which are separated intermediately based on
hydrophobicity.
Isotachophoresis (ITP):
A discontinuous buffer system is used to preconcentrate
injection volumes that are 10 to 100 times the normal. Following a
short focusing period, the sample is separated as in CZE.
References:
- Heegaard, N.H., Robey, F.A. Anal. Chem. 64, 2479 (1992)
- Compton, S.W., Brownlee, R.G. Biotechniques 6, 432-440
(1998)
For more information:
E-mail Kari Hartman ,
CAMIS
or call (603) 862-1696
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