Affinity chromatography is a chromatographic technique in which separation occurs, based on the specific interaction between two molecules. This specific interaction can be between enzyme and substrate, receptor and ligand, protein and nucleic acid etc. affinity chromatography is usually carried out in column which is filled with the supporting material or the matrix. The matrix used should be chemically inert, should exhibit good flow properties and should have sufficient functional groups for the covalent binding of the ligand. The matrix is attached with a spacer arm which is usually made up of CH2 group. The presence of spacer prevents the non-specific interaction of the ligand with the matrix itself. when the mixture of different molecules is passed through the affinity column, only the molecule that have specific affinity for the ligand, binds the lignd. the other molecules are washed away. Next to dissociate the molecule of interest from the ligand, a buffer with different ionic strength of pH is passed. This finally releases the molecule of interest from the ligand and is obtained in the pure form.
30 сен 2024
