Gel-filtration chromatography

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This is also known as Molecular or size exclusion chromatography and uses the principle of column chromatography. The size of the molecules is the basis of separation. The stationary phase is a column consisting of porous beads made up of insoluble but highly hydrated polymers such as polyacrylamide, agarose, biogel [1]. These porous beads have a specific pore size and so are nearly able to separate molecules with a size similar to it [2].

The diameter of the polymers is usually about 0.1mm as such only small molecules could enter these beads. The larger molecules which can’t enter the internal volume of the beads remain in the solution between the beads and so flow more rapidly through the column.

A sample i.e. a protein sample is introduced at the top of the column and eluted with a solvent.

Gel chromatography can separate complex mixtures with great precision and as such is used in areas of biotechnology.

References

  1. Berg J, Tymoczko J and Stryer L. (2007) Biochemistry, 6th edition, New York: WH Freeman.
  2. The Gemini Geek. What is gel chromatography http://www.thegeminigeek.com/what-is-gel-chromatography/. Last accessed on 27/11/2010
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