Chromatography is a most
frequently used laboratory technique for separation and purification of biomolecules
mixture. The mixture is dissolved in the mobile phase (a kind of fluid), which
carries it through the stationary phase (a structure holding another material).
The separation is based on differential partitioning between the mobile and
to the different separation mechanism and principle, chromatography techniques are
classified into different types for separation and purification process of the
various target biomolecules. There are 4 types of commonly used chromatography techniques:
Affinity chromatography, Ion exchange chromatography (IEC), Gel filtration
(also known as Size-exclusion chromatography (SEC) and molecular sieve
chromatography), and Hydrophobic interaction chromatography (HIC).
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chromatography (IEX) is a practical chromatography process that separates ions
and polar molecules based on their respective charged groups. It works on
almost any kind of charged molecule—including large proteins, small
nucleotides, and amino acids.
Gel filtration chromatography, also known as size exclusion chromatography (SEC), is a chromatographic method that separates molecules by their size or molecular weight. SEC can be used for wide ranges of separation and purification of components with large molecular weight differences and low resolution requirement, such as proteins, polysaccharides and other macromolecules.
HIC is a useful method for purification and separation of biomolecules based on their surface hydrophobicity. HIC can be applied in the separation and purification of hydrophobic proteins such as aromatic and aliphatic compounds. In HIC, the matrix material is lightly substituted with hydrophobic groups, such as methyl, ethyl, propyl, octyl, or phenyl groups.