Affinity Chromatography Columns
Affinity chromatography columns separate proteins by providing a stationary phase modified with a ligand that specifically interacts with the protein.
Binding: The target molecule is specifically and reversibly adsorbed by a complementary ligand and immobilized on a matrix. Examples of a complementary ligand include an inhibitor, substrate analog or cofactor, or an antibody which specifically recognizes the target molecule. The selectivity is often based on spatial recognition, a ‘lock-and-key’ mechanism.
Elution: The adsorbed molecule is eluted either by competitive displacement or a conformation change through a shift in pH or ionic strength. Typical molecular pairs are antigens and antibodies, enzymes and coenzymes, and sugars with lectins.