Gel Filtration Chromatography

Gel Filtration Chromatography Background Information

Proteins that differ in mass can be separated by gel filtration chromatography.  In this technique, the column is composed of porous beads made from polyacrylamide, dextran (a bacterial polysaccharide), or agarose (a seaweed derivative).  Proteins flow around the spherical beads in gel filtration chromatography.  However the surface if the beads is punctured by large holes, and proteins will spend some time withing these holes.  Because smaller proteins can penetrate into the beads more easily than larger proteins, they travel through a gel filtration column more slowly than larger proteins.  (In contrast, proteins migrate through the pores in a electrophoretic gel; thus smaller proteins move faster than larger ones.)  The total volume of liquid required to elute a protein from the column depends on its mass; the smaller the mass, the greater the elution volume.  By use of proteins of known mass, the elution volume can be used to estimate the mass of a protein in a mixture.

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