Affinity Chromatography

By

Dr. Mehboob Peeran

Affinity chromatography is a type of adsorption chromatography. In this method the stationary phase or the bed material in the column has biological affinity for the substance to be isolated. The specific properties of the stationary phase are developed by covalently linking an appropriate binding ligand to an insoluble porous support (solid matrix), thus immobilising one of the components of the interacting systems on the solid matrix. The mixture, which contains the specific substance to be separated, is then made to pass through the solid matrix when the ligand, which has specific affinity to the component to be, separated binds selectively to it. The resulting complex is then washed to get rid of other undesirable substances and eluted with a solvent system which breaks up the forces that bind the ligand and the component (desorption), thus making elution of the particular component possible.
These basic steps are illustrated in the following scheme.

Acknowledgement

The author had attended a ten day workshop on the subject conducted by the Primate Research Lab in the Indian Institute of Science, Bangalore, several years ago. Most of the material in this article is from the lectures and information obtained from that workshop