Gel Filtration Chromatography, Molecular Exclusion Chromatography, Molecular Sieve Chromatography, Adsorption Chromatography

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Gel Filtration Chromatography
In this chromatography, various proteins are separated on the basis of differences in their molecular size. This technique of gel filtration chromatography is also known as molecular exclusion chromatography or molecular sieve chromatography.

The basic arrangement for gel filtration chromatography is similar to that for adsorption chromatography. A column made up of glass or steel is taken and packed with a gel.

An Arrangement for Gel Filtration Chromatography to Purify an Enzyme

The most commonly used gel- is sephadex, which is a cross-linked dextran produced by certain strains of bacteria. Several types of sephadex, namely G-10, G-30, G-50, G-100, G 150. G-200, etc. are available, which differ according to their pore size.

When a mixture of enzymes or proteins is poured on top of the column different proteins move downwards according to their molecular sizes and come out from the column in order of decreasing sizes; the larger molecules are eluted first. The elution volume is logarithmically proportional to the molecular size.

Gel filtration chromatography can be used for determining the molecular weight of the protein by calibrating the column with proteins of known molecular weights.

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